Microscopy 
Reeves HAND-BOOK 
OF 
MEDICAL MICROSCOPY. 
REEVES. A HAND-BOOK 
OF 
MEDICAL MICROSCOPY 
FOR 
STUDENTS AND GENERAL PRACTITIONERS, 
INCLUDING CHAPTERS ON 
BACTERIOLOGY, NEOPLASMS, AND URINARY 
EXAMINATIONS. 
BY 
JAMES E. REEVES, M.D., 
to 
MEMBER OF THE ASSOCIATION OF AMERICAN PHYSICIANS; EX-PRESIDENT OF THE 
AMERICAN PUBLIC HEALTH ASSOCIATION; EX-MEMBER OF THE STATE BOARD 
OF HEALTH OF WEST VIRGINIA, AND SECRETARY OF THE BOARD ; EX-PRESI- 
DENT OF THE STATE MEDICAL SOCIETY OF WEST VIRGINIA; CORRES- 
PONDING MEMBER OF THE PATHOLOGICAL SOCIETY OF PHILA- 
DELPHIA; HONORARY MEMBER OF THE CONNECTICUT MEDI- Nl (} 
CAL SOCIETY; AUTHOR OF A PRACTICAL 
ON ENTERIC OR TYPHOID FEVER," ETC., ETC.*/w£CO> Q 
WITH A GLOSSARY V^LlBF 
AND NUMEROUS ILLUSTRATIONS (PARTLY IN COLORS). 
PHILADELPHIA: 
P. BLAKISTON, SON & CO., 
IOI2 WALNUT STREET. 
1894. Copyright, 1894, by F. M. Reeves. 
PRESS OF WM F. FELL & CO. 
1220-24 SANSOM STREET, 
PHILADELPHIA. TO 
MRS. FRANCES M. REEVES, 
MY BELOVED WIFE 
AND 
ONLY COMPANION IN MICROSCOPY, 
WHOSE GREAT MIND AND LOVING HEART HAVE ENCOURAGED 
MY PROFESSIONAL ENERGIES AT EVERY STEP, AND TO 
WHOM I AM INDEBTED FOR THE GREATEST 
HAPPINESS OF MY LIFE, 
THIS VOLUME 
(fSrattfuIIj anti JHost Effectionattlj ©cMcatcir 
BY 
THE AUTHOR. PREFACE. 
This little volume has been written from the stand- 
point of a general practitioner of more than forty years’ 
active experience who thinks he is well acquainted with 
the needs of his professional brethren. 
It cannot fail to strike all those who have paid any 
attention to modern medical education that the micro- 
scope has contributed in very large measure to bring 
about the high scientific tone which characterizes the 
profession of to-day. Indeed, as an aid to correct 
diagnosis, it is the most important instrument in the 
hands of the physician ; yet those who have neglected 
to make themselves familiar with its advantages in the 
daily routine of ordinary practice are in overwhelming 
majority,—the excuses offered for such' delinquency 
being “ lack of time,” “ the expense of a microscope,” 
and “no private teacher,” or “guide-book worthy of 
the name.” 
It is the object of this volume to deny all such ex- 
cuses by showing how small the necessary expense of a 
microscopic outfit—how easy and delightful the way 
to successful work, or for the physician to become his 
own microscopist. 
VII VIII 
PREFACE. 
The book itself proves what may be accomplished by 
singleness of purpose without a teacher, also the truth 
of the old adage, “ Where there is a will there is a 
way.” In other words, I have tried in a plain and 
simple manner to put myself “in his place,” and given 
just such a guide as I felt the need of when I began the 
study of microscopy, and would most cheerfully have 
paid for the instruction a hundred times the price of 
this volume. 
For kindly and valuable assistance in carrying the 
work through the press, I owe many grateful thanks to 
Professor Charles S. Dolley, Professor James Tyson, 
and Dr. John H. Packard. Also, I am indebted to the 
publishers for their painstaking to produce one of the 
handsomest volumes ever issued by them. 
I have no apologies to offer in presenting the work ; 
for it is intended to be useful, and must stand or fall 
by its own measure. 
James E. Reeves. 
Chattanooga, July, 18Q4. TABLE OF CONTENTS. 
I. Introduction:— PAGE 
Importance of the Microscope in Medicine—Neces- 
sary Qualifications of a Physician—Illustrative Cases, . . 17-33 
II. Necessary Microscopic Outfit:— 
Microscope Stands—Eye-Pieces—Objectives—Linear 
Magnifying Powers of Objectives and Eye-Pieces—Illu- 
mination— Achromatic Condenser—Eye-piece Micrometer 
—Double Nose Piece, 34-53 
III. How to Work :— 
Works to be Consulted—Typical Slides—Stock of 
Needed Materials—Importance of Pure Anilin Stains — 
Clean Slides and Covers 54_6i 
IV. Preparation of Animal Tissues:— 
Fixing and Hardening—Effete Methods—Importance 
of a Sharp Knife in Dividing fresh Material—Softening 
Hard Material in Salt-Solution—The Water-Bath—Clear- 
ing and Imbedding—The Paraffin Method—Gas-pipe 
Moulds and Wooden Blocks, 61-68 
V. Recapitulation :— 
Fixing and Hardening of Tissues—Embedding—Mak- 
ing the Cast, 68, 69 
VI. Cutting Sections :— 
The Necessity of a Microtome—Good American Pat- 
terns—Sharpening the Knife—Section Flattener, .... 7°“74 
VII. Fixing Sections Immediately on the Slide :— 
Egg Albumin as a Fixative—Centering the Section— 
Soaking in Turpentine—Washing in Benzol and Alcohol, 74"77 
VIII. Staining and Finishing : — 
Borax-carmine—How to Use the Stain.—Hrematoxy- 
lin Double Stain Based on Delafield’s Preparation.—How 
to Use the Double Stain, 77-^1 
IX. Formulary:— 
The Anilin Dyes—Gentian Violet, Fuchsin, Methyl- 
Violet, Methyl-Blue, Bismarck Brown or Vesuvin.—(No. 1) 
Carbolized Solution Methyl-Blue.—(No. 2) Aqueous Solu- 
IX X 
TABLE OF CONTENTS. 
tion Methyl-Blue.—( No. 3 ) Koch’s Solution Methyl- PAgb 
Blue.—(No. 4) Loffler’s Solution Methyl-Blue.—(No. 5) 
Chenzynski’s Solution.—(No. 6) Plehn’s Solution.—(No. 
7) Unna’s Borax Methyl-Blue.—(No. 8) Unna’s Stain for 
Microorganisms in the skin.—(No. 9) Noniewicz-Loffler- 
Unna Method.—(No. 10) Anilin Oil Methyl-Blue.—(No. 
11) Ehrlich’s Solution Gentian Violet.— (No. 12) Aqueous 
Solution.—(No. 13) Carbolized Fuchsin, Neelsen’s.— 
(No. 14) Aqueous Solution of Fuchsin.—(No. 15) Gibbes’ 
Double Stain.—(No. 16) Carbolized Double Stain.—(No. 
17) Koch’s Solution Methyl-Violet.—(No. 18) Carbolized 
Methyl-Violet.—(No. 19) Bismarck Brown, Concentrated 
Solution.—(No. 20) Carbolized Aqueous Solution.—(No. 
21) Borax-Carmine.—(No. 22) Haematoxylin Double- 
Stain.—(No. 23) Weigert’s Fibrin Stain.—(No. 24) The 
Author’s Method.—(No. 25) Cram’s Solution.—(No. 26) 
Gram’s Method Modified.— (No. 27) Anilin Oil Solution. 
—(No. 28) Anilin Water.—(No. 29) Flemming’s Solution. 
—(No. 30) Ehrlich-Biondi Triple Stain.—(No. 31) Ma- 
lachowski’s Stain.—(No. 32) Toison’s Staining Fluid, . . 81-88 
X. Bacteriology:— 
The Germ-Theory—Classification.—Asiatic Cholera 
—Methods of Artificial Culture—How to make a Plate- 
Culture—Cover-glass Demonstration.—Bacillus of Cholera 
nostras—resemblance to the Comma Bacillus of Asiatic 
Cholera.—Enteric or Typhoid Fever—Diagnostic Char- 
acters and Causal Relation of the “ Typhoid Bacillus ”— 
Cover-glass Preparations and Section-Mounts—The 
Author’s Method of Treating Sections.—Pulmonary 
Tuberculosis—The Importance of the Tubercle Bacillus 
in Diagnosis—Culture-Media—Cover-glass Preparations of 
Sputum, and Method of demonstrating the microorganism 
in tissue.—Croupous Pneumonia—The Micrococcus 
pasteuri of Sternberg, and the Pneumobacillus of Fried- 
laender—Methods of Staining and Differentiation.—Diph- 
theria—The Klebs-Loffler Bacillus—Methods of Stain- 
ing.—Scarlet Fever—The Specific Germ not yet dis- 
covered.—Gonorrhoea—Neisser’s Gonococcus—Failure 
of Cultivation Experiments—Cover-glass Preparations.— 
Syphilis—The Alleged Bacillus of Syphilis—Danger of 
confounding the Smegma Bacillus—Method of Staining. 
—Glanders—Frequency of Occurrence in the Human TABLE OF CONTENTS. 
XI 
Body—the resemblance of the Specific Microorganism to page 
the Tubercle Bacillus—Its Infective Power—Culture Media 
—Cover-glass and Tissue Preparations.—Leprosy—The 
Bacillus a slender rod resembling the Tubercle Bacillus— 
Cultivation experiments not generally successful—Very 
sensitive to some of the Basic Anilin Stains—May be 
demonstrated with the Carbolized Double Stain (No. 16). 
—Tetanus—The Specific Microorganism a “ pin-shape” 
rod having a spore at one end—Discovered by Nicolaier 
—Probably an Infectious Disease—Very responsive to the 
basic Anilin Colors.—Influenza—The alleged Influenza 
Bacillus the smallest of the rod bacteria—May be culti- 
vated in Glycerin-Agar, and stained with (No. 5) Chen- 
zynski’s Solution.—Anthrax—The specific bacillus meets 
every requirement to prove its causal relation—May be 
demonstrated according to Gram’s Method.—Amoeba 
coli—A Protozoon of much clinical interest because of its 
connection with Dysentery and Abscess of the Liver— 
May be stained with Loffler’s Solution (No. 4).—Plas- 
modium malariae—the particular Parasite—Different 
phases of the Hematozoon, intra- and extra-corpuscular— 
Exhibits Ameboid Movement—Pigmented Bodies—Cul- 
ture experiments not successful—For immediate diagnosis, 
the Method recommended by Prof. Jaksch.—Actinomy- 
cosis—A disease of wide distribution in Cattle—the 
fungous character of the disease—Communicable by acci- 
dental inoculation of the fungus to human beings—classed 
with the polymorphic fission-fungi—Artificial cultures suc- 
cessful in various media—Cover-glass and Section pre- 
parations may be made by several Methods, 89-142 
XI. The Examination of Neoplasms:— 
Importance of Clinical Microscopy—The frequency of 
Tissue examinations—A Tumor in histo-pathological sense 
—The different forms—Fibromata, Lipomata, Chondro- 
mata, Osteomata, Papillomata, Adenomata, Lymphomata, 
Myomata, Neuromata, Angeiomata, Lymphangeiomata.— 
The Sarcomata—Fleshy tumors following the Modified 
type of the Connective tissues of the embryo—Large and 
Small Spindle-celled growths—Large and Small Round- 
celled Sarcomas — The Giant-celled Alveolar, Mixed- 
celled and Melanotic Sarcomas—Presenting all degrees 
of Malignancy.—Glioma and Teratoma.—The Car- XII 
TARLE OF CONTENTS. 
cinomata—Structure follows the type of the Epithelial page 
Tissues—Hard and Soft Cancers—Flat-celled and Cylin- 
der-celled Epithelioma—Colloid Cancer—Melanotic Can- 
cer—Mucous Cancer—The Parasitic Theory of Cancers 
and Sarcomas—Methods of Demonstration, 142-162 
XII. Urinary Examinations:— 
How to Collect a Specimen—Composition of Normal 
Urine—Color and Pathological Conditions — Specific 
Gravity—Reaction—How to Handle a Specimen.—Or- 
ganized Constituents — Epithelial Cells — Mucus—Red 
Blood Corpuscles—Leukocytes.—Casts—“ False Casts ”— 
True Casts—Blood Casts—Epithelial Casts—Hyaline Casts 
—Waxy Casts—Granular Casts—Fatty Casts.—Relation 
of Albumin and Casts to Bright’s Disease.—The Surprises 
in Clinical Medicine.— Urea.— Spermatozoa.— Tubercle 
Bacillus and other organisms.—Amorphous Urates—Uric 
Acid—Oxalate of Lime—Triple Phosphates—Ammonio- 
Magnesium Phosphates—Urates of Ammonium and Sodi 
um—Sulphate of Calcium.—Chylous Urine.—Cystin.— 
Hippuric Acid.—Xanthin.—Tyrosin and Leucin.—Cho- 
lesterin.—Summary, 163-200 
XIII. The Blood :— 
Volume and Composition—Red Blood, White Blood, 
and Blood-Tablets—Hayem’s Fixing Medium—The Dis- 
eases of the Blood—Instruments for Counting the Red 
and White Blood Cells—Application of the Thoma and 
Zeiss Method—Toison’s Staining fluid.—Test for Hsemin 
Crystals.—Poikilocytosis.—Animal Parasites 201-210 
XIV. The Sputum :— 
Tubercle Bacilli and other Microorganisms—Methods 
of Technique.—Diphtheritic Membrane and other Exu- 
dates.—Crystalline Forms in Phthisis and Pneumonia.— 
Cholesterin Crystals and Crystals of the Fatty Acids:— 
Lime Crystals, and Concretions, 210-213 
XV. Fecal Discharges: — 
Average Daily Quantity of Feces—Composition— 
Protozoa and Vermes, 213-217 
XVI. Mucous Cylinders, 217 
XVII. Micro-organisms of the Skin, 217 
XVIII. Micro-organisms in Foods and Drinks, . . 218 
Glossary, 221-234 
Index 235-237 LIST OF ILLUSTRATIONS. 
FIG. PAGE 
1. Bausch & Lomb’s Continental Microscope, 39 
2. Zentmayer’s American-Continental Stand, 41 
3. Huyghenian Eye pieces, 42 
4. Achromatic Condenser, 52 
5. Double Nose-piece, 53 
6. Fibres of Linen, Silk, Cotton, and Wool, 57 
7. Dr. Reeves’ Water-bath and Oven, 66 
8. Bausch & Lomb’s Laboratory Microtome, 71 
9. Zentmayer’s “ Ryder Automatic ” Microtome, 73 
10. Centering Diagram, 75 
xi. Forms of Bacteria. {From Schenk), 92 
12. Koch’s Plating Apparatus for “ Setting” Agar or Gelatin, . 98 
13. Platinum Needles, Straight, Hooked, and Looped, .... 99 
14. Damp Chamber for Plate Cultivations, 100 
15. Comma Bacilli of Asiatic Cholera. (Jaksch). [Colored), 101 
16. Finkler-Prior Bacilli. [Jaksch). [Colored), 102 
17. Bacilli of Typhoid Fever; 2 cuts to show both rods and 
“spider-cells.” [Colored), 104 
18. Tubercle Bacilli in Sputum. [Jaksch). [Colored), . . .112 
19. Recent Croupous Pneumonia, 118 
20. Pneumonococci. [Jaksch). [Colored), 119 
21. Bacilli of Diphtheria. [Gould). [Colored), 120 
22. Gonococci Contained in Pus-cells. [Friedlaender). (Col- 
ored), 122 
23. Bacilli of Syphilis. [Gould). [Colored), 124 
XIII XIV 
LIST OF ILLUSTRATIONS. 
FIG. PAGE 
24. Bacilli of Glanders; large and small amplification. (Jaksc/i). 
(Colored), 125 
25. Bacilli of Leprosy. (Coplin and Gould). (Colored), . .126 
26. Bacillus Tetani. (Coplin). (Colored), 127 
27. Bacilli of Influenza. (Gould). (Colored), 128 
28. Bacillus Anthracis. (Coplin). (Colored), 129 
29. Bacillus of Symptomatic Anthrax, Flagellate Form. 
(Gould). (Colored), 129 
30. Amoeba Coli. (Colored), 130 
31. Plasmodium Malariae; pigmented and non-pigmented 
forms, 134 
32. Semilunar Bodies—Extra Corpuscular, 135 
33. Rosette Forms, , 136 
34. Rosette Forms with Segmentation, 136 
35. Spirillum of Relapsing Fever. (Coplin). (Colored), . . 138 
36. Actinomyces—Ray Fungus. (Coplin). (Colored), . . . 139 
37. Osteoid Chondroma. (Gould), 146 
38. Leiomyoma of the Uterus. (Gould), 149 
39. Round-celled Sarcoma. (Gould), 152 
40. Giant-, and Small Spindle-celled Sarcomg. (Gould), . . 153 
41. Melanotic Alveolar Sarcoma of the Skin. (Gould), , . .154 
42. Scirrhus Carcinoma and Medullary Carcinoma. (Gould), 157 
43. Cylindrical Epithelioma, 159 
44. Colloid or Myxomatous Cancer. (Gould), 160 
45. Urinometer, 167 
46. Epithelium, Renal, Bladder, Vaginal, 170 
47. Blood Corpuscles, Changes in. (Landois), 173 
48. Pus-cells in Urine. (Ralfe), 175 
49. Deposit in Acid Fermentation of Urine, 180 
50. Deposit in Ammoniacal Urine, 180 
51. Micrococcus Urese, 183 
52. Fungi in Uringe. (Landois), 183 
53. Graduated Ureameter, 185 
54. Renal Tube Casts. (Landois). (Colored), 187 LIST OF ILLUSTRATIONS. 
JUG. PACK 
55. Normal Semen, 188 
56. Forms of Uric Acid Crystals. {Landois). {Colored), . .191 
57. Oxalate of Lime, 192 
58. Forms of Crystals, Ammonio - Magnesium Phosphate. 
{Tyson), 194 
59. Stellate and Feathery Crystals of Triple Phosphate. ( Tyson), 195 
60. Acid Ammonium Urate. (Landois), 196 
61. Crystals of Cystin, Oxalate of Lime, etc., 197 
62. Hippuric Acid. {Landois), 198 
63. Tyrosin and Leucin, 199 
64. Cholesterin Plates. {Landois), 200 
65. Blood Corpuscles and Blood Plates. {Jaksch). {Colored), 202 
66. Thoma-Zeiss Capillary Tube. {Jaksch), 204 
67. Counting-slide, 205 
68. Ilemin Crystals. {Landois). {Colored), 208 
69. Leukemic Blood. {Landois). {Colored), 208 
70. Blood in Poikilocytosis. {Colored), 209 
71. Filaria Sanguinis Hominis. {Leuckart), 209 
72. Bilharzia Hematobia. {Leuckart), 210 
73. Microscopic Characters of Mixed Saliva. {Landois). {Col- 
ored), 211 
74. Charcot-Leyden Crystals. {Landois), 212 
75. Cephalic end of Tenia Solium; Measly Pork. {Leuckart) 
{Colored), 214 
76. Taenia Saginata. {Leuckart), 215 
77. Trichina Spiralis. {Leuckart). {Colored), 216 
XV HAND-BOOK 
OF 
MEDICAL MICROSCOPY. 
I. INTRODUCTION. 
IMPORTANCE OF THE MICROSCOPE IN MEDI- 
CINE. 
For the purposes of this volume, it would be a waste 
of words and space to repeat the oft-written history of 
the Microscope, and tell how from the beginning with 
a simple convex lens, or magnifying glass, the instru- 
ment has reached marvelous perfection, optical and 
mechanical, and become a necessity in the households 
of Science and Art. 
Neither is it necessary to attempt a recital of the 
many ways in which the microscope has advanced the 
science and art of medicine. Open any book we may, 
and behold the revelations of the microscope. Indeed, 
but for the priceless aid afforded by this instrument in 
the discovery of exact knowledge of the minute struc- 
ture of the human body in healthy and diseased condi- 
17 18 
MEDICAL MICROSCOPY. 
tions, the complete text-books of to-day on anatomy, 
physiology, pathology, bacteriology, and the theory 
and practice of medicine would have been an impossi- 
bility. 
Especially may we boast of the truly wonderful ad- 
vances made in bacteriology—the newest branch in the 
cycle of the medical sciences,—and claim for its all-ab- 
sorbing importance the crowning glory of microscopic 
triumph. By its searching power and incomparable 
penetration into the boundless world of life which lies 
beyond the ken of normal vision, the microscope has 
wrought a complete transformation of once dominant 
ideas concerning the nature and treatment of epidemic 
and contagious diseases; thus dethroning and demolish- 
ing long-cherished idols, whose dicta had not only 
swayed the multitude of medical opinion, but really 
made it hazardous to disregard their precepts in the 
diagnosis and treatment of many diseases. 
In those days, however, the laborers in the medical 
vineyard saw “ through a glass darkly, but now face 
to face;” and in the near future it is confidently 
hoped the Ark of the Covenant with biologic science, 
quickened by the possibilities of microscopic technique, 
will move forward another day’s journey to possess 
richer fields and broader highways in the realms of 
“Biologos.” Then, looking down from some Pisgah 
height, peering deeper and deeper into the little lump 
of protoplasmic jelly, the basis of life, from which the 
cell-wall is elaborated and all living structure springs— 
“ things that visive organs reach not,”—we may, per- IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
19 
chance, find the secret of life and save the children of 
men from “ the pestilence that walketh in darkness and 
the destruction that wasteth at noonday.” 
The time has now come when all progressive physi- 
cians and surgeons, general practitioners and specialists 
alike, must either themselves possess sufficient skill in 
microscopic technique for the faithful and proper dis- 
charge of the high obligation which rests upon them in 
the diagnosis and treatment of diseases, or else be able 
to command the ready service of some accomplished 
microscopist and pathologist to do such necessary work 
for them. In no other way can they conscientiously 
perform their duty to those whose lives are placed in 
their hands, and fully meet all the reasonable require- 
ments of advancing medical science. 
To be a learned, capable physician in the full sense 
requires that he shall bean all-round doctor,—physicist, 
chemist, anatomist, physiologist, pathologist, bacteri- 
ologist, and microscopist: have all these varied qualifi- 
cations correlated and blended with his professional 
name and life-work, never forgetting for a moment that 
it is the welfare of the patient he should always have 
first at heart. 
Pathologic microscopy is the key to open up the way 
to rational and successful treatment. It is the founda- 
tion to all therapeutic deduction ; and without its pro- 
per study and acquaintance, the Practice of Medicine 
and Surgery would be but little better than guesswork 
and a dangerous play with human life. 
How should the astronomer hope to be able to com- 20 
MEDICAL MICROSCOPY. 
prehend his subject without having a thorough know- 
ledge of mathematics and skill in the manipulation of 
the telescope ? or the pilot on an ocean steamer find 
his way to the port for which he set sail without a com- 
pass to guide his course across the trackless waters? 
Such, precisely, is the relation which pathologic mi- 
croscopy occupies to the practice of medicine in all its 
branches; it touches every department of the pro- 
fession with immense practicality, and nothing can 
supplement its power in the search for abstract truth 
concerning diseased conditions and their proper treat- 
ment. 
To comprehend disease at the bedside or wherever else 
it is confronted, the physician must have in his mind’s 
eye a clear and exact picture of the changes going on in 
the minute structure of the part involved ; he must know 
the nature of the disease—“hear the cry of the suffering 
organ,” and distinguish it—before he can reasonably 
hope to repair the wrong. It is not enough merely to 
look into a microscope. A person who cannot tell a 
microscope from a telescope may do as much, and go 
away from the instrument without having derived the 
least profit by his curiosity. 
There is a vast difference between seeing and observ- 
ing. To be able to observe correctly requires not only 
eyes to see, but fingers to feel and a heart to sympathize. 
Every avenue of the mind and soul should be engaged 
and cultivated to the most delicate impressions— 
indeed, all these being necessary qualities of prepara- 
tion, which neither books alone nor the possession of the IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
21 
most complete microscopic outfit, or both combined, 
can supply. 
In pathology, as elsewhere in the domain of science, 
there is no short road to knowledge and technical skill. 
The eye, the hand, and the mind must all alike be 
trained by long use before a technique most complex 
and difficult can be mastered and scientific research 
undertaken with hope of credit. 
When I began the use of the microscope, nearly 
twenty years ago, there were but few pathologic 
laboratories in the United States, and even the chairs 
of Histology and Pathology were “more honored in 
the breach than the observance” in at least nine- 
tenths of the first-class medical colleges. But now, 
what a happy advance. In every large city well 
equipped pathologic laboratories have sprung up to 
meet the professional demand; every first-class medical 
college can now boast of its laboratory equipments and 
“show its faith by its works; ” and thus in all direc- 
tions we may discover the march of scientific medicine. 
At the beginning of my experience with the micro- 
scope, I fully realized the fact that the instrument was 
more ornamental than useful in my hands, but I never 
became discouraged by my failures to meet my own ex- 
pectations ; and so, on and on, I have kept going to 
this day, when I find myself more than ever before in 
love with my silent but most instructive companion,— 
the wonderful instrument which has aided me in a 
thousand ways of doing good to my patients by a 
correct understanding of their diseases. 22 
MEDICAL MICROSCOPY. 
The whole world of science was at once quickened 
into activity by the discoveries of Pasteur, Koch, and 
others in bacteriology; and nothing of all the revela- 
tions of the microscope is or can be more interesting to 
the medical scientist, or of greater importance to man- 
kind, than the knowledge thus derived concerning living 
microorganisms,—their almost increditable minuteness ; 
some of them rods, others spirals, others chain-like, 
others ovals, others spheres, each class arranging itself 
in its own peculiar order; their wonderful life-histories 
and marvelous rapidity of growth, either by division 
or spores, some well-known forms doubling their 
number every hour, so that a single bacterium may 
produce eight million, three hundred and eighty-eight 
thousand, four hundred and eight of its kind in the 
space of twenty-four hours, each variety elaborating its 
own specific poison in the genesis of disease, and of 
incomparable activity in the processes of fermentation 
and putrefaction; some of them having power of motion 
by means of flagella at one or both ends ; either aerobic 
or anaerobic, and producing pigments of various colors ; 
liquefying gelatin or not; cultivated in various media 
outside the animal body, and having a thermal death- 
point ranging from 1220 to 2120 F., according to the 
particular variety, culture-medium, and time of ex- 
posure: a temperature of 2120 F., maintained for five 
minutes, being sufficient to kill all pathogenic organ- 
isms and their spores. 
But notwithstanding the value of the microscope as an 
aid in advancing pathology and bacteriology, it is not to IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
23 
be supposed that sharp cultivation of the senses by clinical 
experience is less necessary now than before the instru- 
ment became of such high importance in all the branches 
of the practice of medicine and surgery. To feel the 
pulse, look at the tongue, hear the heart’s beat and other 
physical sounds in the chest, and go over the patient 
from head to foot to find out the diseased organ or fail- 
ure of function is just as necessary to-day as before in- 
struments for diagnostic precision were used. The 
symptoms of pulmonary involvement by disease are not 
less important because we have the pleximeter and 
stethoscope to discover physical signs, and the micro- 
scope with which to examine the sputum. And should 
the surgeon be excused for less skill with the knife 
because he cannot tell certainly a sarcoma from a carci- 
noma, or a benign tumor from a malignant growth with- 
out the aid of the microscope ? 
While the science and art of medicine have been 
greatly improved in modern times by the proper use of 
ingenious instruments, clinical skill is none the less im- 
portant for the correct interpretation of symptoms and 
the causes of diseases. Indeed, by reason of the now 
indispensable instrumental aids in the hands of the 
physician, clinical knowledge and skill are more neces- 
sary than ever before. In tracing disease to its origin 
the necessary methods are far more complex and intri- 
cate than those hitherto employed. The high aim now 
is at the precision of the exact sciences, and in that 
way pathology and therapeutics are both aided and 
advanced. 24 
MEDICAL MICROSCOPY. 
The following examples, selected from a large number 
that could be furnished, are given in proof of the every- 
day value and necessity of the microscope in the prac- 
tice of medicine and surgery : — 
Case I.—A pale-faced young woman, unmarried, came to me on 
account of her uneasy sensations, from which she had suffered, more 
or less, for the previous six months. She had a dragging gait, and 
complained of loss of appetite, pain in the back and weakness in 
the lower limbs, palpitation of the heart, sleeplessness, nervousness, 
“ a ball in the throat,” and a constant feeling of general weariness. 
She was dark under the eyes, had lost flesh, suffered from leucor- 
rhea, and was plainly hysterical. 
After hearing the history, I told her to bring me for three succes- 
sive days a morning and evening specimen of her urine. The next 
day she came with the first bottle. The urine was pale and cloudy, 
of low specific gravity, and slightly alkaline. Microscopic exam- 
ination of the deposit revealed numerous spermatozoids; and thus 
the cause of all her ailments was most surprisingly discovered. 
Case II.—A married man, having a creamy-looking discharge 
from the urethra and painful urination, came to me with a sad 
story—sad because he charged his wife with infidelity and blamed 
her for his condition, firmly believing he had gonorrhea and that he 
contracted the disease from her. To make very sure, however, that 
there should be no mistake about the case before beginning divorce 
proceedings and making public his shame, he determined to have 
me make a microscopic examination of the running matter from 
his privy member. 
The absence of Neisser’s gonococcus in the pus-discharge was my 
warrant for telling the unhappy man that he had charged his wife 
wrongfully. She had an acrid leucorrhea, and he, as a consequence 
of his wife’s innocent affliction, had a non-specific urethritis— 
“ merely this and nothing more.” He went away from my office 
wiser and happier than when he came, and was soon well of his 
“ bad disorder.” IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
25 
Case III.—A female child, three years old, suffered from frequent 
and painful micturition. The vulva was inflamed and much swol- 
len, accompanied with copious leucorrheal discharge. This condi- 
tion of the symptoms had lasted eight or ten days when a specimen 
of the child’s urine was brought to me for examination with the re- 
port that the mother had tried various domestic remedies, including 
castile soap washes, but without effect. A drop of the deposit in 
the urinary specimen placed under the microscope showed large 
crystals of ammoniaco-magnesium phosphate and a female oxyuris, 
or seat-worm. Of course, after the discovery of the worm, the 
cause of the little patient’s distress was made plain and the curative 
treatment at once instituted. 
Case IV.—A physician in Western New York sent me what he 
supposed to be “ an undescribed intestinal parasite,” and with it the 
following history of the case :— 
The patient, a female, aged thirty-six, and unmarried. She was a 
chronic sufferer from “ nervous dyspepsia,” and every few days 
passed strange-looking masses from the bowels, like the specimen I 
had received. With each like discharge the patient became more 
and more alarmed about her condition, and the case was a puzzle 
to the attending physician. 
After examining the specimen I wrote the physician and asked 
him if his patient was not fond of oranges? The hint was enough, 
and he replied, “ Yes, sometimes she devours a dozen a day.” The 
supposed intestinal parasite was nothing more than a portion of the 
undigested pulp of an orange. 
Case V.—The patient had suffered from sore and painful ears for 
several weeks. The external meatus was the seat of trouble, and 
besides great tenderness and pain, there was a slight creamy dis- 
charge. A cover-glass preparation of the matter showed the Asper- 
gillus nigricans, or fungous character of the affection. Thorough 
douching with a solution of the hyposulphite of soda, and powdered 
alum blown into the ears, cured the disease promptly. 
Case VI.—An unmarried female, aged twenty-seven, a seam- 
stress, enjoying the benefits of a comfortable home, consulted me on 26 
MEDICAL MICROSCOPY. 
account of her suffering for eighteen months from nausea and vomit- 
ing, coming on within an hour after each meal, regardless both of 
quality and quantity of food taken into the stomach. She had lost 
flesh, and was pale and feeble in appearance ; but her will-power 
never failed, and kept her from giving up her trade-work. 
For relief from her distressing condition she had been under treat- 
ment by several physicians, and, from first to last, had taken every 
dose—pharmaceutic and domestic—she could hear of as ever 
having been of service in dyspepsia. Besides medicinal doses and 
the observance of dietetic rules, she had had her stomach washed 
out several times, but all of no lasting benefit. 
I placed a fragment of the yeast-like vomited matter on a slip of 
glass and added to it a drop of liquor potassa colored with me- 
thylin blue. When placed under the microscope, the well-known 
fungus, Sarcina ventriculi, was easily discovered as the probable 
cause of the stomach trouble. 
A dose composed of one drop of carbolic acid and ten grains of 
hyposulphite of soda, administered before each meal, did more good 
than all else that had been given; but I soon lost sight of the case 
and do not know whether the patient was permanently benefited by 
the treatment or not. 
Case YII.—Miss , aged twenty-six, of healthy parentage, 
average weight 130 pounds, dark hair and eyes, bright and cheerful 
disposition, with active home-life and good hygienic surroundings. 
Nearly four years previous to the recent illness below described 
she had an attack of dysentery which lasted two weeks before con- 
valescence was fully established. After recovery from that sickness 
she grew fat and hearty, and remained the very picture of perfect 
health until July, 1892, when she began to droop ; lost her appetite, 
had a hacking cough, was feverish in the afternoon and chilly in 
the morning, with constipated bowels and sleeplessness. Such 
was her condition for ten or twelve days, during which she lost flesh 
and strength in a marked degree. Then suddenly followed exacer- 
bation of all the features of the case, and the attending physician 
was hurriedly summoned. There had been a severe chill, and the IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
27 
pain in the right chest, extending from the axilla to the liver, was 
now so acute that she could not take a full breath without a moan. 
The cough was most harassing and unappeasable, expectoration 
copious and of pneumonic appearance, respiration hurried and 
jerking, temperature and rapid pulse. 
Day after day this condition continued with but little change, ex- 
cept that the expectoration became more and more copious. The 
persistent cough, heavy expectoration, night-sweats, wasting of flesh, 
and progressive debility led the family and friends to fear the case 
was one of “ hasty consumption,” and to clear up the diagnosis a 
specimen sputum was sent to me for microscopic examination, by 
the attending physician. 
The absence of tubercle bacilli and presence of pus cells, frag- 
ments of hepatic tissue, and liver-cells, was unmistakable proof that 
the case was one of hepatic abscess communicating with a bronchus 
certainly, and, probably, also the colon. 
After two months’ confinement from this serious illness, during 
which she several times passed by stool pus-like matter, the recovery 
was sufficiently complete to enable her to go out-of-doors and visit 
among her friends. She kept up for several months and then was 
taken down again to endure the same suffering as at first. During 
this second illness, the expectoration sometimes showed, mixed with 
the pus-discharge, little gelatinous masses streaked with blood, which 
I examined for Amoeba coli with negative result. The patient is 
now, to all appearances, perfectly well. 
Case VIII.—A babe six weeks old that had an easy birth, and 
thrived well at its mother’s breast for the first fifteen or twenty days, 
was attacked with diarrhea and soon transformed from a fine, plump, 
healthy infant into a starved nursling with sunken jaws and wrinkled 
skin, notwithstanding an undiminished supply of mother’s milk. A 
specimen of the secretion was brought to me for microscopical ex- 
amination and found to contain colostrum corpuscles and compara- 
tively few milk globules. The presence of colostrum corpuscles, 
normally present during the first eight or ten days after childbirth, 
was proof of the poor quality of the milk. The infant was taken 28 
MEDICAL MICROSCOPY. 
away from the breast, fed artificially, and at once commenced to im- 
prove. 
Case IX.—A young man discovered a small tumor, not larger 
than a filbert, under the skin a little below and to the left of the 
navel, and tender on pressure. It soon began to increase in size, 
and he went to a surgeon, who advised its immediate removal and 
performed the operation. The specimen was brought to me for 
microscopic examination, and turned out to be a small round-celled 
sarcoma. My diagnosis was received with evident distrust by the 
surgeon, he thinking the growth nothing more than “ a simple fibrous 
tumor.” I replied, “ I hope so.” 
Within the next eighteen months quite a number of like tumors 
made their appearance under the skin in different parts,—principally 
in the groins and about the root of the neck,—and at the expiration 
of two years from the date of removal of the first tumor the patient 
was dead,—the death-certificate giving correctly the cause of death, 
“ Multiple Sarcoma.” 
Case X.—A clerk in the office of the W. & A. Railway, at Atlanta, 
came home to Chattanooga on account of his sickness, which had 
lasted for several days and presented symptoms pointing to an attack 
of “ the fever” which then prevailed in that city. He had started 
out with a decided chill, head-ache, pain in the back, and soreness 
of the muscles from head to foot, loss of appetite, thirst, nausea, 
with occasional vomiting, and inability to sleep. 
When the patient came under my observation his condition for 
the first three days so closely resembled genuine enteric or typhoid 
fever, with which I had been so long familiar, that I thought the op- 
portunity was at hand of showing my professional neighbors the 
wide difference in the mode of access, march, and complications 
between this case and the usual clinical history of the malarial fever- 
cases met with at all seasons in this community and the country 
round about. 
There was, however, something about the case when the symp- 
toms were all critically studied, after a few days’ attendance, that 
excited my uncertainty of the diagnosis, and I appealed to the mi- IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
29 
croscope for a decision. A drop of blood taken from the finger 
showed the presence of both crescents and pigment granules, thus 
proving the malarial character of the case. 
The temperature range was more fitful than in typhoid fever, per- 
haps, for one day running at in the morning and 102° in 
the evening; the next day, 103° in the morning and ioi° in the 
evening. On the tenth day the afternoon temperature reached 
the following day it did not go above At no time 
was diarrhea present. Within two weeks the patient was dis- 
charged.* 
Case XI.—Mrs. , aged forty-three, the mother of five 
* It was not until after my removal to Chattanooga, in 1888, 
that I had frequent opportunity of seeing this hydra headed form of 
continued fever,—variously called “ typhoid fever,” “typho-mala- 
rial fever,” “ simple malarial fever,” “ continued malarial fever,” 
etc.,—such as the case above reported. I can remember several 
occasions when I was in doubt, during the first week of the case, 
because of the fitful and irregular march of the symptoms. 
The poison of malaria and the contagium of enteric or typhoid 
fever are not antagonistic forces, and may conjointly display their 
effects, thus producing a commingling of symptoms truly confusing 
to the clinician; but such a blending of types cannot give origin to 
a distinct, permanent type of fever. Such products are hybrids in 
the full sense of the word, and wholly incapable of multiplication. 
When the enteric or typhoid element is conjoined, then that is the 
only quantity which is capable of propagation, or spreading from 
person to person. 
Enteric or typhoid fever is communicable from person to person, 
and cannot spring up de novo. In its prevalence, it may assume dif- 
ferent degrees of severity,—appear either in isolated cases with but 
feeble contagious power, or seize whole households, attack whole 
neighborhoods, or spread over large districts, according to the pre- 
vailing “ epidemic constitution.” 
Such is not the history of the outbreak and spread of continued 30 
MEDICAL MICROSCOPY. 
children, short in stature, accustomed weight one hundred and 
fifty pounds, ruddy complexion, rheumatic or gouty tendency, and 
enjoying a full share of home comforts, including the protection 
afforded by good hygienic surroundings, while warm and per- 
spiring, exposed herself to a cool draught in an open porch, 
where she sat for a few minutes “ to cool off.” She soon became 
chilly, and from that moment her illness began. For the next week 
she complained of stiffness and soreness of the limbs ; had head- 
ache, backache, and was more or less feverish and thirsty. At 
the end of that time her attending physician pronounced the case 
enteric or “ typhoid fever,” and treated it accordingly, he being 
very careful to avoid laxative medicine. 
On account of rapid deepening of all the symptoms, I saw 
malarial fever. The clinical features are also markedly different. 
The temperature-curve is more variable and uncertain as to the time 
of rise and fall; the remissions are more regular and decided; the 
gastro-hepatic involvement is more pronounced ; and the absence of 
diarrhea, epistaxis, tympanites, rose-colored eruption, dull and list- 
less expression of countenance, and great prostration of muscular 
strength, that mark enteric or typhoid fever. Besides, the duration 
of malarial fever is shorter. 
In enteric or typhoid fever one attack affords immunity, usually, 
for the remainder of life. This is as constant as the safety from a 
second attack of measles, smallpox, scarlet fever, or yellow fever. 
In malarial fever the patient is more liable to a second attack than if 
he had not had the disease, and so on his liability increases with each 
successive attack. 
After convalescence from enteric or typhoid fever, there is in most 
cases rapid accumulation of flesh which is out of all proportion to 
the muscular strength ; the hair falls out, and in due time the person 
is either greatly improved in general health or quickly succumbs to 
tubercular consumption, if he have such tendency. After convales- 
cence from malarial fever, the subject is for months in poor health 
and strength, his countenance is sallow, and digestion feeble. IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
31 
the patient in consultation on the tenth day after the date of ex- 
posure on the “ porch ” where she was said to have taken cold. 
She had fever, cough, with frothy expectoration streaked with 
blood, hurried and jerking respiration, dusky countenance, and 
slightly contracted pupils; puffiness of the eyelids, feet, and 
ankles; twitching of the muscles, itching of the skin, bowels not 
moved for twelve hours, urine scanty and dark-colored, and, withal, 
great drowsiness of manner. 
An immediate examination was made of the urine. It was of 
acid reaction with sp. gr. 1012, and highly albuminous. Under 
the microscope the deposit revealed all sorts of casts—blood casts, 
epithelial casts, both pale and dark granular casts, a few hyaline 
casts, and a still fewer number containing oil drops. Besides casts 
there were red blood and white blood corpuscles, crystals of uric 
acid, and granular matter. 
After these discoveries in the urine, the plan of treatment was 
changed so as to include as quickly as possible the revulsive and 
hydragogue effect of a full dose of compound jalap powder, under 
which influence the patient waked up from her uremic intoxication. 
During the next couple of weeks the jalap powder was several 
times repeated. 
To make a long history short, it may be stated that within the 
next three months the patient was able to go out-of-doors, and after 
twelve months she seemed to have so far recovered as to be able to 
resume control of her household affairs ; but all the while she was 
extremely sensitive to changes of weather, and suffered from slightly 
swollen feet and ankles. Now, ten years afterward, notwithstand- 
ing the reminder of a little swelling of the feet and ankles, she is 
still in the enjoyment of a fair state of health by avoiding, in her 
manner of living, excesses and exposures of every kind. 
In the diagnosis and treatment of urinary and renal diseases, I 
am so very dependent upon the microscope for guidance that I 
should be compelled to decline the medical management of all such 
cases if from any cause I were deprived the use of the instrument, 
for without it my treatment would be but guesswork. 32 
MEDICAL MICROSCOPY. 
The mere discovery of albumin in the urine is not alone sufficient 
to establish the diagnosis of Bright’s disease. Indeed, there may 
be a form of this malady with but slight, if any, trace of albumin, 
especially in the interstitial variety of the disease so common in 
whisky drinkers or habitual “ tipplers ” and hard-pressed business 
men, in which cases there may be no complaint of serious trouble in 
the kidneys until the subject is struck down with a convulsion. 
Any life insurance agent or other person totally ignorant of the 
pathology and treatment of Bright’s disease may learn in a half- 
hour’s time how to make the test for albumin ; but the burden of most 
important knowledge concerning the condition of the kidneys—im- 
portant alike to the company and the applicant—can only be ob- 
tained in the field of the microscope by a competent manipulator 
and appreciative observer. 
Case XII.—Mrs. , aged thirty-one, of tuberculous family, but 
enjoying accustomed good health, took cold from wet feet, and 
sickened with symptoms so closely conforming to the usual clinical 
history of enteric or typhoid fever that no doubt existed in the mind 
of her experienced physician concerning the correctness of his diag- 
nosis—namely, “ typhoid fever; ” and the case was treated accord- 
ingly for more than three weeks. After the beginning of the third 
week there was but little change in the symptoms, nothing to mark 
particular attention except the increased amount of heavy expectora- 
tion and an occasional night-sweat. Cough and bronchial rales were 
prominent features among the prodromata, and the nose bled once 
or twice. From the beginning of the illness there was a tendency to 
looseness of the bowels. The rose-colored spots were not searched 
for because of the assumed certainty of diagnosis. The anticipated 
and anxiously looked for critical days came and passed without sign 
of improvement, and in that way the fifth week was reached without 
abatement of any of the symptoms. All the while, the appetite was 
poor and capricious, sometimes amounting to entire disgust of food. 
On account of the lingering character of the case—the unappeas- 
able cough and copious expectoration, wasting of flesh and strength, 
easily relaxed bowels, hectic and night sweats—it was deemed ad- IMPORTANCE OF THE MICROSCOPE IN MEDICINE. 
33 
visable to have a microscopic examination made of the sputum, 
when lo! swarms of tubercle bacilli were discovered. The patient 
died within four months from the beginning of her illness, the death- 
return being “ Acute Tubercular Phthisis ”—to which might have 
been added, of the typhoid type. 
And thus I might go on narrating case after case in 
my experience to prove the value of the microscope in 
the practice of medicine and in the pursuit of scientific 
truth. 
The microscope is the watchful sentinel to warn us 
against the enemies of our flesh. It scans with search- 
ing eye the food we eat, the water we drink, the cloth- 
ing we wear, and the very air we breathe. And this is 
not all of its usefulness. Besides helping us to under- 
stand the nature of our diseases and the way of their 
proper treatment, it is the unerring detective of the 
cheat and adulterator of either food or medicine, and 
dreaded alike by the forger and the murderer. As such, 
the instrument is a blessing to all mankind. 
Already the literature of the microscope is volumin- 
ous, and thousands of active workers are in the field 
gathering the ripe wheat of the harvest, yet there is room 
for a more general knowledge of the best and simplest 
methods of technique. To aid in filling such need, and 
encourage the use of the instrument in the hands of 
medical students and physicians, by simplifying its use, 
is the end whereunto, and with many sincere wishes 
for its success, this little volume is sent to the medical 
profession. 34 
MEDICAL MICROSCOPY. 
II. NECESSARY MICROSCOPIC 
OUTFIT. 
The difficulties which I myself encountered when I 
took up the study of microscopy without a teacher— 
especially my innocent ignorance of just what was really 
necessary in a microscopic outfit for a practising physi- 
cian—may now be turned to the profit of those of my 
readers whose limited means forbid unnecessary ex- 
pense in the purchase of a microscope. 
The splendid catalogues that are now sent out by 
microscope makers and dealers, while marking the 
wonderful advancement that has been made in micro- 
scopic science, are not only misleading, but truly be- 
wildering to the beginner at the tube by their attract- 
ive show of illustrations and explanatory descriptions. 
In proof, see the variety of microscope stands, different 
grades of objectives, and all sorts of accessories, includ- 
ing microtomes of ingenious design, also photographic 
apparatus, and a hundred other useful things worth 
having by those who can afford them ; in other words, 
the offer of “complete ” outfits varying in price from 
$40.00 to $1200.00. 
It.was in such uncertainty of my actual needs that I 
spent many hard-earned dollars unnecessarily, and to- 
day can produce enough old keepsakes from such pur- 
chases to buy—if I had the money they cost—a suffi- 
cient microscopic outfit for a beginner. 
In the selection of a stand there are really but four 
important points necessary to be considered, namely, NECESSARY MICROSCOPIC OUTFIT. 
35 
solidity, steadiness, fine adjustment by micrometer 
screw, and plenty of room with all needful fittings 
for sub-stage condenser and other accessories. 
It must not be supposed that the larger and more 
ornamental the stand the more powerful the microscope. 
A twenty-five-cent chromo may be set in a hundred- 
dollar frame without in the least improving the quality 
of the picture, but a hundred-dollar picture may be put 
in a twenty-five-cent frame without detracting from the 
value of the piece. If the picture is good, we are 
wholly indifferent as to the quality of the framework 
which holds the canvas. And so with the choice of 
a stand. It is the eye-piece at one end of the tube and 
the objective at the other which make the microscope, 
and the cost of the objective may be much or little, 
according to the wish of the purchaser to accommodate 
his needs. 
Fortunately, in these days of Columbia’s lead in the 
arts and sciences, we no longer have to go abroad for 
our microscopes and other scientific instruments, nor 
for instruction how to use them. Our own microscope 
makers are now not only fully able to supply the home 
demand, but have gone into foreign markets with their 
superior instruments and challenged comparison with 
the very best patterns in England, France, and Ger- 
many. In fact, everything that can be needed or de- 
sired by the working microscopist may be obtained in 
this country, and at reasonable price. 
Owing to the great variety of microscopes in the 
market,—foreign as well as home products,—the in- 36 
MEDICAL MICROSCOPY. 
experienced person in the technique of the instrument 
should, if possible, get some trustworthy expert to assist 
him in making the choice, for unless such precaution 
be taken to prevent mistake, nine chances to one the 
purchaser will sooner or later regret his outlay as so 
much money unwisely spent, if not, as well, thrown 
away. 
A very good microscope fitted with all needful acces- 
sories for medical and ordinary bacteriologic work 
may be had for the very low price of $50.00 or $60.00, 
and a very complete outfit—such as will answer the 
physician for a lifetime—can be purchased for $100.00, 
or less, from either of our first-class makers and 
dealers. 
It is not too much to say that Bausch & Lomb, 
Rochester, N. Y., and Joseph Zentmayer, Philadelphia, 
are probably the largest manufacturers of microscopes 
in this country; and an instrument purchased from 
either house may be confidently relied upon to be just 
what is claimed for it in the catalogue description. 
Besides these two well-known makers, there are others 
who turn out perhaps equally good work, but have not, 
as manufacturers, engaged such long general public at- 
tention concerning the superiority of their wares. 
Chicago is well represented by W. H. Bulloch and the 
McIntosh Co. ; Rochester, N. Y., besides Bausch & 
Lomb, the Gundlach Optical Co. ; and Philadelphia 
can boast also of the house of J. W. Queen & Co., 
together with the agency of R. & J. Beck. It should 
also be stated that in supplying the market with micro- NECESSARY MICROSCOPIC OUTFIT. 
37 
scopes there are several justly distinguished specialists 
who confine their skill to the manufacture of objectives. 
A notable example is the H. R. Spencer Co., whose 
high-power lenses are truly marvelous in performance. 
Then, again, there are dealers who carry, besides some 
particular microscope with their own trade-name, various 
stands and objectives, both new and second-hand, and 
at such an establishment sometimes a great bargin can 
be secured. The society-screw and standard length of 
tube make it possible to use all sorts of objectives with 
any make of stand, and thus this branch of the micro- 
scope business is more and more encouraged by persons 
in search of cheap microscopes. But it should not be 
forgotten by beginners in microscopy that much risk is 
run in purchasing a second-hand outfit, no matter how 
reputable the maker, without the approval of some 
expert to determine its true value. A very prevalent 
impression with the uninitiated is that the larger and 
more ornamental the stand, the greater the magnifying 
power of the glass, or that it must be a very fine instru- 
ment because it “magnifies seven hundred times,” as 
a physician once said in praise of his microscope, and 
to prove his skill in detecting tube-casts in a case of 
Bright’s disease. 
My own equipment consists of a Bausch & Lomb 
professional stand with Abbe condenser, and the follow- 
ing battery of objectives: ii in., T\, 1, -fa, and y the 
last two being oil-immersion objectives, and all of them 
“ first-class.” The j1 is a Spencer, and for excellence 
in performance is unsurpassed by any glass of like power. 38 
MEDICAL MICROSCOPY. 
This outfit leaves nothing better, in my judgment, 
to be desired. The Bausch & Lomb T\* and are 
dry, and so useful in every-day work that I should not 
know how to get along without them. Because of its 
wide angle and freedom from the inconvenience of im- 
mersion fluids in the examination of fresh mounts, the 
i is well adapted to ordinary bacteriologic work, and 
is indeed a jewel of incomparable value to any working 
microscopist. 
Besides these old friends of mine, there are other well' 
known lenses, whose fortunate owners praise them in 
the highest for superior performance under the most 
severe tests. In this class I am pleased to speak of the 
Gundlach objectives, also of the specialty of Queen & 
Co., their 1-15 oil-immersion objective, which has won 
testimonials from various experts whose good opinion 
in such matters is worth having. 
My advice to the beginner is to start with first-class 
lenses, no matter how cheap the stand ; but the stand 
must be fitted with a sub-stage condenser, to get the 
full performance of the objective and do satisfactory 
work in bacteriology. 
The accompanying illustrations show the most inex- 
pensive as well as the most useful patterns of stands. 
For all ordinary use, the cheapest stand will answer 
as well as the costliest; and to that end the Continen- 
tal, shown on the opposite page, is good enough for 
* This superb glass, the T40-tk, Messrs. Bausch & Lomb do not 
keep in stock, and can only furnish it on special order. Fig. i. 
(Cut one-half of actual size.) 
Bausch & Lomb Continental Microscope. 
39 40 
MEDICAL MICROSCOPY. 
anybody. It is a happy combination of the useful with 
the ornamental, but not really better than other stands 
shown in various illustrated catalogues. The selec- 
tion—when all of the patterns are so good—is more a 
matter of individual taste than exercise of judgment. 
The particular machine we are most familiar with we 
like best, and recommend it for that reason to our 
friends—“ merely this and nothing more.” 
But the selection of a microscope and all needful 
accessories is one thing ; the successful use of the instru- 
ment another and quite a different performance. Be- 
fore the instrument can be properly manipulated and its 
fields correctly interpreted, the student must by long and 
patient effort bring his senses up to the full requirement 
of cultivation ; he must understand what is meant by 
accurate microscopic vision, and become master of a 
most difficult technique before expecting to reap the 
plenteous harvest which lies within reach of industrious 
effort and singleness of purpose. 
By failing to appreciate the necessity of such prepara- 
tion, many a physician after purchasing a microscopic 
outfit has learned to his sorrow that it was of no immedi- 
ate use to him. As well might the purchaser of a grand 
piano expect on going to the key-board for the first time 
to be able to strike harmonious chords and bring forth 
sweet sounds without previous and patient training of 
his senses to the most delicate impressions, as for the 
physician to expect to use a microscope successfully 
without first learning how to see and observe. For 
this reason, the show of a microscope in a physician’s 
office is often more ornamental than useful. NECESSARY MICROSCOPIC OUTFIT. 
Fig. 2. 
41 
Zentmayer American-Continental Stand. 42 
MEDICAL MICROSCOPY. 
Every microscope when it comes from the maker is 
supplied with one or more eye-pieces, the Huyghenian 
or negative, which consist of a longer or shorter tube 
with a plano-convex lens at either end, the curved or 
convex surfaces looking downward. 
EYE-PIECES AND OBJECTIVES. 
Fig. 3. 
Huyghenian Eye-pieces,—C and D 
The shorter or more shallow the tube of the eye-piece, 
the greater the magnifying power. The upper lens is 
known as the eye-glass, the lower one the field-glass. 
The accompanying full-sized sections of Huyghenian 
eye-pieces show the depth and construction of oculars 
C and D. The A has a depth of 2 in., B in., C 1 EYE-PIECES AND OBJECTIVES. 
43 
in., D y in., and E y in. It should be observed 
that the shorter the tube of the eye-piece the deeper 
the curvature or convexity of the lower surfaces, and 
that, correspondingly, the diaphragm is more contracted 
and the aperture in the cap of the eye-glass smaller. 
Unlike the fixed measure of the society-screw, with 
which all first-class objectives are now made, there is no 
uniformity of diameter of eye-pieces from different mi- 
croscope makers, and hence the necessity of buying all 
extras from the maker of the stand. 
If the eye-piece is a good one, it correctly amplifies 
the image made by the objective; and so, if the objec- 
tive is imperfect in any respect and gives a distorted 
image, the eye-piece will magnify it in proportion to 
the depth of the tube. 
Very strong eye-pieces—C, D, and E—are really of 
but little use, except to try the excellence or poverty 
of an objective which, if well made as to its optical pro- 
portions, will bear stronger eye-pieces than one of faulty 
construction. The good rule constantly gaining favor 
is to work with a low eye-piece and rely upon the ob- 
jective for necessary amplification. 
A perfect eye-piece is free from all cloudiness, striae, 
spots, or other defects ; and the field of vision should 
be sharp and intensely black. Specks of dust or other 
dirty accumulations may be easily seen by rotating the 
eye-piece. All such foreign matter should be carefully 
removed by blowing the breath on the glass and wiping 
with an old silk handkerchief or piece of well-washed 
chamois. 44 
MEDICAL MICROSCOPY. 
Beginners at the monocular microscope find it diffi- 
cult, with both eyes open, to look into the tube with one 
eye and see nothing whatever with the other eye. To 
accommodate those who cannot accustom themselves to 
work with both eyes open, various devices to blind one 
eye have been employed ; but a simple piece of dark 
blue blotting paper or card-board, with a hole through 
the center to admit the cap of the eye-glass, will answer 
as well as anything else that has been offered. 
Objectives.—Each maker of objectives makes claim 
of some private formula for working his combinations 
of different qualities of glass—crown and flint—to suit 
varying density and refractive indices, also in grinding 
the curves of his lenses and spacing the different systems 
in the mounting. Upon such assumed correct knowl- 
edge of optical principles and claim of faultless tech- 
nique he bases the superiority of his instruments; but, 
unfortunately, there is no established or acknowledged 
standard by which to decide such claims and fix their 
relative merits. Every observer, like the maker of his 
objective, is a law unto himself and an unknown, be- 
cause unstable, quantity. There are, indeed, so many 
accidental circumstances, both of person and environ- 
ment, which may influence one’s judgment of the per- 
formance of any particular objective that very often a 
distinction is made without a difference. The present 
condition of health of the operator, whether his nervous 
system has been impressed or disturbed by excessive 
indulgence of any kind, his stomach full or empty, 
digestion good or bad, also the state of the weather and EYE-PIECES AND OBJECTIVES. 
45 
the quality of light—all these,—are active factors in 
determining the appreciable performance of an objec- 
tive. In other words, every expert microscopist is aware 
of the fact that in many cases—especially when using 
high powers and in the study of bacteria—the objective 
is blamed when the fault lies in the observer himself. 
There are, however, certain easily discovered quali- 
ties by which the value of an objective may be deter- 
mined. These are : defining power, flatness of field, 
penetrating power, and resolving power. 
Defining power depends upon the correction for 
spherical and chromatic aberration. Until within the 
last few years complete achromatism was believed to be 
impossible on account of the disproportionate dispersion 
at different parts of the spectrum. But by* the genius 
of Prof. Abbe and his distinguished co-workers in 
optical science—Dr. Schott and Carl Zeiss—suppression 
of the secondary spectrum was triumphantly accom- 
plished, and since then chromatic and spherical aberra- 
tion have been practically eliminated. 
If an objective is not properly corrected for spherical 
aberration the lines of objects appear cloudy and indis- 
tinct, truly lacking in defining power. For discovery 
of such defect, a stage micrometer will afford a conven- 
ient test—to see if the lines are clear and sharp, like the 
lines in a copper or steel plate. If not corrected also 
for chromatic aberration, colored fringes are in the 
field—blue, if the lens be under-corrected ; red, if 
over-corrected. 
Flatness of field gives as clear and perfect definition 46 
MEDICAL MICROSCOPY. 
at the periphery as at the center of the field. Without 
this important quality, the objective is not worth having; 
but the examiner should make sure, if any inequality of 
clearness of field exist, that the fault is in the objective. 
Unless the slide is perfectly flat, a really good lens may 
be rejected as lacking in flatness of field. Here again 
the stage-micrometer may be used in the test; but it 
should be remembered that the ruled lines may not all 
be equally sharp, hence it will be necessary to run them 
in various directions. If the lines are exactly parallel 
to each other, sharp alike at all distances from the 
center as at the center of the field, then the perfection 
of the objective in this particular respect is assured. A 
better test, perhaps, is by comparison of the field under 
a well-known faultless lens of the same power and angle. 
Penetrating power is the depth or measure to which 
an objective can reach into an object or tissue and 
expose the character of its make-up. In histologic 
work good penetrating power is an essential quality of 
an objective. The surface of the field is not only to be 
explored, but the component structures beneath must 
be looked into. The lower the angle of aperture the 
better the penetrating power, is the rule, which is 
equivalent to saying the higher or wider the angle of 
aperture the less its power of penetration. A good 
test-object is a section of human liver, double-stained. 
Defining and resolving power both increase with and 
depend upon the absolute angle of aperture of the 
objective, provided the aberrations are well corrected. 
A glass may have all needful defining power, yet if it EYE-PIECES AND OBJECTIVES. 
47 
have not sufficient aperture its field is accordingly con- 
tracted. In other words, the wider the aperture the 
greater the resolving power, if corrections for aberrations 
are accurate. The larger the window the greater the 
flood of light that enters, and wider the landscape view; 
hence the value of wide angles of aperture in bacterio- 
logic work. 
The working distance of all wide-angle objectives is 
small,—especially with dry objectives,—requiring the 
thinnest of cover-glasses, or No. i. 
In spite of the excellent quality of the objective, the 
thickness of the cover-glass exerts much influence over 
the field, particularly when using high powers. An 
object which under a No. i cover gives a sharp image 
becomes blurred and dimmed when viewed under a No. 
2 cover. To overcome the influence of varying thick- 
ness of cover-glasses, lenses are made with correction 
collar, by means of which the relative positions of the 
individual systems of lenses may be changed to suit the 
thickness of the cover-glass; but it requires a practised 
eye at the tube to find the exact turn of the screw which 
stops at the best effect. Collar-correction seems to be 
more necessary in dry than immersion lenses. 
The usual tests for resolving power of high-angle 
objectives are the frustrates of certain diatoms, the 
Amphipleurapellucida being the most difficult to resolve. 
But this test is too high for a dry lens, unless it be one 
having the widest possible angle of aperture. 
Immersion Objectives have certain great advan- 
tages over the dry system. They afford larger working- 
distance than is possible with a dry lens of the same 48 
MEDICAL MICROSCOPY. 
angle; and the denser the immersion fluid the greater the 
working-distance. The drop of liquid—water, glycerin, 
cedar oil, bisulphid of carbon, or other medium—which 
connects the front lens and the upper surface of the 
cover-glass serves the important purpose of partially ex- 
tinguishing the two glass surfaces, and thus prevents 
loss of light, which always takes place from air surfaces. 
By the immersion system, more rays of light pass into 
the microscope, the medium approximating the same 
effect as an enlargement of the angle of aperture. 
Immersion objectives do not require the same preci- 
sion of adjustment of collar-correction for thickness of 
cover-glass that is so necessary in the use of dry lenses. 
The discovery of this important effect led to the con- 
struction of non-adjusting immersion lenses, which are 
easily used, very satisfactory in performance, and have 
become great favorites with all classes of microscopists. 
To the busy physician they are a great help, for besides 
more rapid work they require less skill to use them. 
The latest triumph of Mr. Edward Bausch, of the firm 
of Bausch & Lomb, Rochester, N. Y., in the production 
of his non-adjustable 1-12 inch oil-immersion objec- 
tive, is something of which all American microscopists 
may justly feel proud ; for the performance of this re- 
markable low-priced lens leaves nothing to be desired by 
the working histologist and bacteriologist. 
The instructions given on a preceding page for the 
care of eye-pieces will, as far as they go, equally apply 
to the proper care of objectives. The beginner should 
be content to use low and medium powers in order 
thereby to prepare himself for more difficult work. At EYE-PIECES AND OBJECTIVES. 
49 
least, he should not resort to the use of a high-power 
objective until he has learned how to handle such a 
glass safely and properly. The lack of such knowledge 
and caution has been the ruin of countless valuable 
objectives, not to mention the smashing of cover-glasses 
also, by unskilful persons. 
Even in the most expert hands the front glass becomes 
now and then soiled with balsam, but no damage is 
thereby done unless too much benzol or other cleansing 
agent is used. Two or three folds of old cotton or silk 
should be placed over the end of the finger, slightly 
moistened with benzol, and then whipped back and 
forth two or three times over the face of the glass. 
After that, the breath should be blown upon it and 
then gently wiped again. 
LINEAR MAGNIFYING POWERS OF OBJECTIVES AND 
EYEPIECES.. 
Objectives. 
55 
ro 
55 
55 
-IN 
5 
z 
«w 
z 
-IN 
z 
z 
-w 
z 
f 2 in. (A) 
| ij in. (B) 
eye-pieces: \ i in. (C) 
1 fin- (D) 
l i in. (E) 
16 
21 
3° 
42 
60 
25 
35 
5° 
70 
100 
33 
42 
66 
84 
132 
40 
55 
80 
110 
160 
50 
70 
loo 
140 
200 
70 
102 
140 
205 
280 
176 
247 
353 
495 
7°5 
210 
295 
420 
590 
84O 
265 
385 
530 
770 
1060 
Tube Length 8.5 
IN. = 
2l6 MM. 
Tube Length of 160 mm. 
Objectives. 
Z 
z 
z 
z 
z 
z 
z 
z 
H® 
Hm 
N 
Nlrt 
-w 
-ICC 
M 
f2 in. (A) 
360 
45° 
570 
69O ' 
18 
37 
196 
267 
422 
1 in. (B) 
54° 
660 
845 
1025 1 
26 
52 
285 
400 
626 
eye-pieces: 1 in. (C) 
720 
900 
1140 
1380! 
37 
74 
392 
534 
844 
1 2 in. (D) 
1080 
*32° 
1690 
2050 
52 
104 
570 
800 
1252 
i 5 m. (E) 
1440 
1800 
2280 
2760 
74 
148 
784 
1068 
1688 
Calculated for a Tube Length of 8.5 in. — 216 mm. 50 
MEDICAL MICROSCOPY. 
INSTRUCTIONS FOR USING THE EYE-PIECE 
MICROMETER. 
As the eye-piece micrometer is not compared directly 
with the object itself, but only with the image of it 
formed in the focus of the eye-piece, it is only when the 
exact proportion between the size of the object and 
that of its image is known that measurements of the 
object can be readily determined by the eye-piece 
micrometer. 
This proportion depends upon : ist, the focus of the 
objective ; 2d, the distance of the image from the ob- 
ject ; 3d, the focus and the place of the field-lens when 
the latter is situated between the objective and the image. 
As these relative conditions are not of equal value in 
all microscopes, and consequently the relative size of 
the images, as formed by different objectives of the 
same rating, are not always the same, these sizes have to 
be ascertained for each microscope and objective sep- 
arately. 
To reach this result, a reliable stage micrometer 
should be used as an object, and its image accurately 
measured with the eye-piece micrometer. 
The figures designate the proportion of the linear 
measure of the object, the latter being taken as 1. 
The actual size of any object is, therefore, obtained 
when the size of its image, which is obtained by direct 
measurement by means of the eye-piece micrometer, is 
divided by the figures so obtained. 
For adjustable objectives, the figures are intended for 
close adjustment. ILLUMINATION AND ACCESSORIES. 
51 
In instruments having draw-tube, make measurements 
when tube is at standard length. 
25.4 mm. = 1 inch. 
The proper use of the mirror is one of the most im- 
portant parts of microscopic technique ; yet it is often 
least studied and therefore undervalued by the operator. 
All first-class stands are provided with a double mir- 
ror,—one side plane, the other concave. The mirror 
serves to reflect the light on the object to be examined. 
In ordinary use, the quality is known as transmitted or 
central light. The concave surface reflects the incident 
rays in a convergent direction to the well-hole in the 
stage, and should be used with high-power objectives. 
The plane mirror gives less intense illumination, and 
is suitable for use with low powers. 
The best light is day-light when the sky is clear and 
bright. But direct rays from the sun should be avoided 
as a rule. In sky-light the rays are all practically paral- 
lel, while those from a lamp or other artificial source 
converge strongly from the luminous center. 
In contradistinction with transmitted or central 
light, the more intense and searching power of oblique 
light becomes sometimes a necessity. For this use, the 
mirror should have all needful swinging motion in order 
that it may be placed in the required lateral position. 
This quality of light—either from the direct rays of the 
sun or from an artifical source—is necessary in resolv- 
ILLUMINATION AND ACCESSORIES. 52 
MEDICAL MICROSCOPY. 
ing various test-objects; but great skill in manipulation 
is required to reach the best effect of the mirror. 
When choice of position can be had, the microscope 
should face a north window to receive the very best 
quality of day-light. Working in strong light, either 
natural or artificial, endangers the eyesight, and should 
be avoided. In all my twenty years’ experience with 
the microscope I have not spent, I am sure, altogether a 
week’s time by lamp-light ; and for that reason can now 
boast of unimpaired eyesight. 
The Achromatic Condenser is a very necessary 
Fig. 4, 
Achromatic Condenser. 
aid of high-power objectives, in order to concentrate 
the light to a greater extent than can possibly be done 
with the mirror alone. In bacteriologic investigations, 
especially, the condenser cannot be dispensed with ; but 
in its use care must be taken to insure the coincidence 
of the optic axis with that of the objective. 
Corresponding with oil-immersion objectives we have 
now also oil-immersion condensers, some of them able 
to utilize the rays of 130° balsam angle. All of the 
best patterns are provided with various diaphragms and ILLUMINATION AND ACCESSORIES. 
53 
a blue-glass plate or disc. The “ Abbe Condenser ” is 
the type of the best, but there are others and less ex- 
pensive ones that will answer every purpose. The con- 
denser manufactured by Messrs. Bausch & Lomb is one 
of these. Its numerical aperture is sufficient—about 
1.42—to accommodate and aid the performance of ob- 
jectives of the widest angle. It may be worked both 
dry and immersion. 
A microscope without a sub-stage condenser of some 
sort is of but little use to the physician. 
Fig. 5. 
Double Nose-piece. 
While the Double Nose-piece is a great time- 
saving convenience in changing quickly from one 
objective to another, it is a dangerous instrument; for 
unless very great care is exercised in its use the front 
glass of the objective may be struck in making the 
sweep, and ruined. 
A nose-piece out of “center” is worse than a 
nuisance ; hence the necessity of its thorough testing 
before fitting it permanently to the microscope. If cor- 
rectly fitted, this accessory is worth ten times its cost. 54 
MEDICAL MICROSCOPY. 
III. HOW TO WORK. 
And now, having purchased a good microscope, with 
all needful accessories, the student is ready to begin work. 
The first thing necessary is a strong work-table of the 
ordinary height, and placed, if possible, in front of a 
north window. The feet of the table should be shod 
with thick rubber to prevent jarring from passing vehi- 
cles on the paved street. The top should be a slab of 
plate-glass overlying several thicknesses of common, 
white, unglazed paper, such as newspaper printers use. 
The glass-plate may be secured at any glazier’s shop at 
cheap price, because of the broken pieces always on 
hand from broken windows and doors of business houses. 
To aid him in successful work, the student should 
supply his library as soon as possible with all needful 
books covering the departments of Microscopy, His- 
tology, Pathology, and Bacteriology. The complete 
list of such publications is already large—too large 
for reproduction in these pages. 
There will be no difficulty in making a selection from 
the following-named authors, whose distinguished labors 
have greatly advanced microscopy in particular and 
medicine in general:—■ 
Bausch, Edward. Manipulation of the Microscope. New edi- 
tion. Bound in cloth. 
Carpenter, W. B. The Microscope and its Revelations. Seventh 
edition ; revised, enlarged, and partly rewritten by Rev. W. H. 
IN MICROSCOPY. HOW TO WORK. 
55 
Dallinger, F. R. s. Octavo, cloth, 1050 pages, 800 illustrations, 
including a number of lithographs. 
Davis, Geo. E. Practical Microscopy. Cloth, 436 pages, with 
310 illustrations and colored frontispiece. An English work, 
having extensive reference to American instruments, etc. New 
and revised edition. 
Gage, S. H. The Microscope and Microscopical Methods. 
Fourth edition; revised and enlarged. Octavo, 96 pages, illus- 
trated with six plates and eight figures in the text; bound in 
flexible cloth. 
Gould’s New Illustrated Medical Dictionary. 
James, Frank L. Elementary Microscopical Technology; a 
Manual of the Art of Mounting. 107 pages, illustrated; flexi- 
ble cloth. 
Frey, Heinrich. The Microscope and Microscopical Technology. 
Translated and edited by Geo. R. Cutter, m. d. 
Phin, John. Practical Hints on the Selection and Use of the 
Microscope. This is an excellent hand-book that we can highly 
recommend. It also treats briefly, yet practically, on the mount- 
ing of objects. Cloth, extra, 231 pages. New edition, illustrated. 
Wormley, Theodore G. Micro-chemistry of Poisons, including 
their Physiological, Pathological, and Legal Relations; with an 
Appendix on the Detection and Microscopic Discrimination of 
Blood. 
IN HISTOLOGY AND PATHOLOGY. 
Delafield and Prudden. Hand-book Pathological Anatomy and 
Histology. Fourth edition, 715 pages, 300 illustrations. 
Friedlander, Carl. Microscopical Technology. For Use in the 
Investigations of Medicine and Pathological Anatomy. Trans- 
lated from the second enlarged and corrected edition by Stephen 
Yates Howell, M. A., M. D. 
Gibbes, Heneage. Practical Pathology and Morbid Histology. 
Heitzmann, C. Microscopical Morphology of the Animal Body 
in Health and Disease. 
Klein, E. The Elements of Histology. 56 
MEDICAL MICROSCOPY. 
Piersol, Geo. A. Normal Histology. 
Prudden, T. M. A Manual of Practical Normal Histology. 
Schaefer, E. A. The Essentials of Histology. 
Stowell. Microscopical Diagnosis. Octavo, 250 pages, profusely 
illustrated with wood-cuts and lithographed plates. By Chas. H. 
Stowell, M. D., and Louisa R. Stowell, M. s., of the University of 
Michigan. 
Wethered, Frank. Medical Microscopy. 
Woodhead, G. Sims. Practical Pathology. 
Ziegler, E. A Text-Book of Pathological Anatomy and Patho- 
genesis. Translated by D. MacAlister. Three parts complete in 
one volume. 
IN BACTERIOLOGY. 
Abbott, A. C. The Principles of Bacteriology. 
Crookshank, Edw. M. A Manual of Bacteriology. 
Dolley, Chas. S. The Technology of Bacteria Investigation. 
Explicit Directions for the Study of Bacteria, their Culture, 
Staining, Inoculation, Mounting, etc. i2mo., cloth, 263 pages. 
Fraenkel, C. Text-Book of Bacteriology. Translated by J. H. 
Linsley. 
Hueppe, Ferdinand. The Methods of Bacteriological Investi- 
gation. Translated by H. M. Biggs. 8vo, cloth. 
Jaksch, Dr. Rudolph v. Clinical Diagnosis. The Bacteriologi- 
cal, Chemical, and Microscopical Evidence of Disease. 
Klein, E. Microdrganisms and Disease. An Introduction into 
the Study of Specific Microorganisms. 
Kuhne, A. Practical Guide to the Demonstration of Bacteria in 
Animal Tissues. Translated and edited by Dr. V. D. Harris. 
Prudden, T. Mitchell. The Story of the Bacteria and their Re- 
lations to Health and Disease. i6mo, 143 pages, cloth, 1889. 
Salomonsen, C. J. Bacteriological Technology for Physicians. 
Schenk, Dr. S. L. Manual of Bacteriology. 
Sternberg, Geo. M. A Manual of Bacteriology. 
Vaughan and Novy. Ptomains and Leucomains and Bacterial 
Proteids; or, The Chemical Factors in the Causation of Disease. HOW TO WORK. 
57 
Besides guide-books, the beginner should supply him- 
self with a goodly number of typical mounts—histologic, 
Fig. 6. 
Linen Fibers X 150 Diam. 
Silk Fibers X 200 Diam 
Cotton Fibers X zoo Diam. 
Woolen Fibers X 2°o Diam. 
pathologic, and bacteriologic—for study and compari- 
son with his own efforts at mounting tissues and making 58 
MEDICAL MICROSCOPY. 
cover-glass preparations. He should also as soon as 
possible become familiar with all sorts of extraneous 
matters commonly found in the microscopic field,— 
hairs, fibers of wood, cotton, flax, and silk; cat-hairs, 
mouse-hairs, particles of feathers, etc. The various 
kinds of starch granules will afford interesting study ; 
also the non-pathogenic fungi. 
A still further advanced step will be the examination 
of various organized deposits,—the squamous epithelial 
cell; epithelium from the bladder, ureter, and pelvis of 
the kidney,—each variety with its differential shape, and 
both healthy and fatty ; red and white corpuscles, pus- 
cells, casts from the renal tubes—blood-castS, epithelial 
casts, granular casts, fatty casts, and hyaline casts ; cylin- 
ders composed of urates, or false casts; spermatozoids 
and casts from the seminal tubes; fragments of morbid 
growths, mucus and fibrinous threads, expectorated 
matters, etc. All these maybe definitely known within 
a short time after commencing the use of the micro- 
scope, and every step in advance becomes lighter and 
lighter because of the intensely interesting character of 
the subject. 
The necessary stock of materials need not be 
large or expensive, as shown by the following list: — 
One-half gross white glass slides, 3x1, ground edges. 
One-half ounce square cover-glasses, No. 1, assorted 
sizes. 
One curved-pointed forceps. 
One dozen test-tubes, assorted. 
One “ pipettes. HOW TO WORK 
59 
One spirit lamp. 
Litmus papers. 
One quart commercial alcohol. 
One pint Squibb’s absolute alcohol. 
One “ Bullock & Crenshaw’s benzol. 
Four ounces xylol. 
One pint spirits turpentine. 
Two pounds hard paraffin. 
Either a Bausch & Lomb or Ryder microtome. 
One Reeves’ water-bath and oven. 
One ounce Canada balsam and xylol, in drop-bottle. 
Two ounces Delafield’s hematoxylin. 
One ounce pulverized borax. 
One-half ounce “No. 40” carmin. 
One half “ gentian violet (in powder), Griibler’s. 
Four ounces anilin oil. 
One-half ounce methyl violet (in powder), Griibler’s. 
One half “ methyl-blue “ “ 
One-half “ safranin “ “ 
One-half “ Bismarck brown “ “ 
One-half “ rosanilin hydrochlorid (fuchsin),* Grii- 
bler’s. 
* In ordering “ fuchsin ” from a dealer, it should be remem 
bered that there are several salts of rosanilin—the sulphate, hydro- 
chlorate, and acetate. The acetate will not hold its (magenta) color, 
and is entirely worthless for staining tubercle bacilli. The only 
reliable salt is the hydrochlorate, and it should always be so written 
in making an order for “ fuchsin.” The German preparation 
(Griibler’s) is the best in the market (like all other of his anilin 
stains); but I have sometimes been led to think that this distin- 60 
One-half ounce rosanilin acetate, Griibler’s. 
One-quarter ounce sulphindigotate soda. 
One eight- “ graduate measure. 
One one- “ “ “ (c. c. scale). 
One six- “ glass funnel (ribbed). 
One three- “ “ “ 
One one- “ “ “ 
One-half dozen six- or eight-ounce wide-mouth glass- 
stopper bottles. 
One-half dozen two-ounce glass-stopper, narrow-mouth. 
One brass table. 
One one-ounce capped dropping-bottle. 
Half-dozen wood boxes, each to hold 25 slides. 
All of the glassware named in the above list may be 
had at any drug store. I have constantly supplied my- 
self in that way with all needful bottles, finding the 
wide-mouth j4- lb. German chloral bottles ad- 
mirably suited to my wants. 
Clean Slides and Covers.—Slides and cover- 
glasses when first received are more or less greasy and 
dirty, and must therefore be well cleaned before use. 
New glasses may be cleaned by immersing them in 
MEDICAL MICROSCOPY. 
guished Chemist’s name is not always a sufficient guaranty of the 
purity and genuineness of the article it covers. Not long ago I 
purchased in New York city some “ fuchsin ” that was nothing 
more, probably, than magenta of English manufacture. Certainly, 
it could not have come from Griibler’s laboratory. Staining solu- 
tions of all kinds ready made and of the most reliable quality can 
be obtained from Voigt Bros., Chemists and Pharmacists, Chatta- 
nooga, Tenn. PREPARATION OF ANIMAL TISSUES. 
61 
liquor potassa for a moment, then washed thoroughly in 
water, and next wiped perfectly dry before placing them 
in alcohol to be ready for use. 
Old mounts may be quickly taken down by pouring 
over the slides boiling water and letting them stand in 
it five minutes or more. By this means the cover-glasses 
may be slipped off easily, after which slides and covers 
may both be cleaned with benzol, followed with alco- 
hol. In cover-glass preparations, after washing in ben- 
zol, the thin glasses must be dipped in warm water to 
loosen the baked coating, after which the final cleans- 
ing may be accomplished by slipping the glass between 
the folds of a soft cloth held between the thumb and 
forefinger. 
IV. PREPARATION OF ANIMAL 
TISSUES. 
Very little progress, if any, has been made within the 
last eight or ten years in the preparation of tissues for 
microscopic use—fixing and hardening, cutting sections, 
staining, clearing, and mounting,—notwithstanding the 
appearance of numerous hand-books, guides, and other 
special treatises on the subject. Indeed, it is really sur- 
prising that successive authors should have deemed it 
necessary to repeat so many worn-out and useless 
methods : Muller’s Fluid and other chromic compounds 
—which, possibly, may be excused in special cases, 
such as dealing with specimens from the central nervous 
system, but is most damaging to tissues requiring 62 
MEDICAL MICROSCOPY. 
bacteriologic study,—the same method of imbedding, 
and the same old diagram for folding a “card-board 
tray” in which to make a paraffin cast; the same instruc- 
tions for cutting sections and floating them in a beaker 
containing spirits of turpentine; staining them in a 
watch glass; next, their decoloration with acid alcohol 
in another shallow glass; next, their dehydration with 
strong alcohol; then clearing them in oil of cloves; 
and finally, after much unnecessary handling and whip- 
ping of the cell-elements in transferring the sections 
from one re-agent to another, they are “lifted” to the 
slide, where they are variously sealed, ringed, and 
finished. 
It is the aim of this little volume to cut loose entirely 
from all effete methods and lead the beginner to a 
technique so simple, easy and inviting, that it will put 
him heartily in love with his microscope for all the great 
things it will be worth to him in the practice of medi- 
cine and surgery. 
The waste of valuable pathologic material because 
of the generally uncultivated state of the science of 
pathologic histology, not only among the masses of the 
medical profession, but even with those who occupy pro- 
fessorial positions, is greatly to be deplored. But the 
light is now fast breaking in all directions of the medi- 
cal horizon, and soon—may we not hope—it will be high- 
twelve ! 
What rare opportunities are missed in every commu- 
nity for the collection of tissue-material of the most in- 
structive and valuable character, from lack of ability PREPARATION OF ANIMAL TISSUES. 
63 
on the part of the medical observer to see beyond 
the macroscopic appearances? Within the last two 
weeks two such examples have come to my know- 
ledge. In one case, a laparotomy was made to remove 
a supposed ovarian tumor, but an immense kidney was 
found to be the offending tumor. Luckily, before the 
mass became completely spoiled from want of proper 
care, a little specimen was secured, and on microscopic 
examination revealed carcinoma with easily differentiated 
inclusions of so-called “ psorosperms.” 
In another case, of still greater interest, an intra- 
uterine tumor was found post-mortem, and carelessly 
thrown aside as a “ degenerating fibroid.” A small 
portion of this tumor was, by the merest accident, 
placed in my hands, and turned out to be probably an 
unprecedented curiosity in uterine pathology. Besides 
papillomatous and myxomatous appearances in some 
parts, there were fields of cartilage cells, showing, pos- 
sibly, the teratomatous character of the specimen. 
When I asked for more of the material, the answer was, 
it had been “ thrown away.” 
In handling fresh material the importance of a sharp 
knife—as sharp as a barber’s razor—cannot be overesti- 
mated. Hacking and tearing with a dull knife may 
utterly ruin a valuable tissue for microscopic study. 
In collecting portions of brain the necessity of a thin, 
keen blade needs no argument. And at every subsequent 
division the sharper the knife the better the result. 
For killing and hardening of tissues, nothing is better, 
I think, than a one per cent, alcoholic solution of cor- 64 
MEDICAL MICROSCOPY. 
rosive sublimate, twenty volumes of the solution to one 
of tissue. 
After soaking in this solution for twenty-four hours, 
the specimen is hard enough to be cut up into very thin 
slices—say, the one-fourth of an inch thick—and should 
then be transferred to commercial alcohol to remain 
other twenty-four hours ; next, it should be placed into 
absolute alcohol, there to remain not less than twenty- 
four hours, provided there is no hurry for the result of 
the microscopic examination. 
Many specimens of connective tissue become too hard 
after going through alcohols to be cast and cut success- 
fully. In all such cases, particularly in dealing with 
pieces of dense tumors, the material should first be 
softened in a ten per cent, solution of common salt for 
a few hours, then washed and soaked in commercial 
alcohol, then go into benzol, and next into paraffin- 
benzol. 
Tissues containing lime-salts may be successfully de- 
calcified by soaking them for twelve hours or longer, if 
necessary, in a twenty-five per cent, solution of nitric 
acid in seventy per cent, alcohol. When the process of 
decalcification is complete, the specimen should then be 
washed in tap-water, and further disposed of in the or- 
dinary way. 
It is possible to make a permanent tissue-mount within 
the short space of five hours from the time the specimen 
is taken either from a living or dead body. A very 
small, thin piece is placed at once into absolute alcohol, 
to remain one hour; then into benzol for one hour and PREPARATION OF ANIMAL TISSUES. 
65 
a half; then into melted paraffin a half hour or more ; 
then cast, cut, and mounted,—all this within four or five 
hours. 
In ordinary work, and when time is no object, from 
absolute alcohol the specimen is laid between blotting 
papers for a moment that any floating drops of alcohol 
may be absorbed, and then it is ready for more perfect 
dehydration and clearing up in benzol. Here it must 
remain a longer or shorter period, according to the 
density and size of the particular piece. So long as air 
bubbles are imprisoned in the tissue, it is not prepared 
for the next step, or for soaking in a solution of paraffin 
in benzol, liquefying at 90° Fahr. In this mixture— 
kept at the liquefying point—the specimen may soak 
from one hour to twenty-four hours, according to the 
convenience of the manipulator. Unless I happen to 
be in a hurry to make a finished mount, I give the 
specimen an over-night bath, or at least several hours, in 
the paraffin solution. 
The next step is to transfer the specimen to melted 
paraffin, hard or soft, according to the season, winter 
or summer, and kept just at the melting point, from 
one to eight hours, according to size and density of the 
tissue, or until air bubbles cease to rise to the surface 
of the paraffin—like so many glistening little beads 
around the border. 
The accompanying illustration shows a water-bath 
and oven devised by myself and advertised in Bausch & 
Lomb’s catalogue. It is so simple in its construction 
that any tinner can make it. The paraffin cup has a 66 
capacity of about six ounces, contains a lifter and a 
cap, also a thermometer. The water-bath may be placed 
over a small gas-jet, or kerosene lamp flame, supported, 
of course, by a frame or rest of some sort. The ther- 
mometer is the ordinary dairy scale, with registry to 
160° F., and costs but a few cents. The water-bath 
complete, need not cost more than a few dollars. 
MEDICAL MICROSCOPY. 
Fig. 7. 
Water Bath and Oven, Dr. Reeves’, with thermometer. Heavy polished cop- 
per, with paraffin cup and lifter. Size 8% inches high by 10 inches wide, 
outside diameter. 
Embedding.—And now, all things being ready to 
make the paraffin cast, the procedure of embedding is 
as follows : Take an ordinary prescription blank, lay it 
on any smooth surface—preferably, a small slab of plate- 
glass or marble—and pour upon it enough paraffin to 
cover the size of a half dollar; then as quickly as pos- PREPARATION OF ANIMAL TISSUES. 
67 
sible catch up and lift the specimen from the bottom of 
the cup and place it on the film of paraffin, with best 
surface downward; next, place over the specimen thus 
laid a mold sufficiently large to cover it without pinch- 
ing, and then pour the cylinder nearly full of paraffin. 
The cast should be cooled with ice, if in summer time, 
or set out-of-doors if made in winter. When the paraf- 
fin is sufficiently cold and set, the cast is then pushed 
out of the mold with the thumb. If it is inclined to 
stick to the mold, a few taps against the table will 
make it easy to slip. After removal from the mold, 
the cast is cut down from the end at which it was poured 
until the specimen is nearly touched. The disc thus 
made is then held for a moment over the flame of a 
spirit lamp to melt its surface and stick it to a wooden 
block fitted in size to be received in the jaws of the 
microtome. If the cast is perfect, it will not shrink or 
become depressed at the surface of the specimen em- 
bedded. 
Celloidin is employed in preference to paraffin and 
all other media by many reputable authorities and prac- 
tical workers; but after fair trial of both, and long use 
of paraffin for all purposes for which an embedding 
material is used, I like it the better of the two. Per- 
haps, for embedding the globe of an eye celloidin 
may have some advantage; but even this exception 
may be more a matter of habit and favoritism than 
anything else. 
The paraffin method, along with many superior 
claims, I think, over all other media, is free from the 68 
MEDICAL MICROSCOPY. 
objection of holding a part of the staining material, 
particularly when the anilin dyes are used. 
The molds referred to may be had at any plumber’s 
shop. They are simply sections, an inch long and 
from an inch to an inch and a quarter in diameter, of 
gas-casing, made either of iron or brass, and used 
to cover ordinary gas-pipe connecting chandeliers. 
The wooden blocks, on the ends of which to stick the 
casts, may be obtained at any carpenter’s shop. This 
method of holding the cast in the microtome is the very 
best that can be employed. 
V. RECAPITULATION. 
Fixing and Hardening of Tissues:— 
1. A i per cent, alcoholic solution of corrosive sub- 
limate—time, 24 hours: 20 volumes of solution 
to 1 of tissue. 
2. Remove the specimen and cut into thin slices, 
not more than inch thick; then transfer 
to commercial alcohol—time, 24 hours: not less 
than 20 volumes of alcohol to 1 of tissue. 
3. Absolute alcohol—time, 24 hours: 20 volumes to 
1 of the specimen. 
4. The specimen placed between blotting-papers for 
a moment. 
5. Benzol until all imprisoned air bubbles are ex- 
pelled and the tissue thoroughly saturated and 
cleared : 1 hour to 8 hours. RECAPITULATION. 
69 
6. Solution of paraffin in benzol, liquefying at 90° 
Fah.—time, from 1 hour to 24 hours. 
Embedding :— 
7. From No. 6, the specimen is transferred to pure 
paraffin in the cup for that purpose belonging to 
the water-bath and oven, the temperature of the 
melted paraffin never to exceed 140° Fah.—time, 
from 1 hour to 8 hours, or until air bubbles cease 
to rise to the surface. 
Making the Cast:— 
8. Place a piece of writing paper, or prescription 
blank, on any smooth hard surface and pour 
upon it a thin coating of paraffin. 
9. Lift the tissue-specimen from the melted paraffin 
in the cup and place it on the coated paper. 
10. Set over the specimen the mold and pour it 
nearly full of the hot paraffin. 
it. Cool quickly as possible, and free the cast from 
the mold by tapping the cylinder several times 
on the table and pushing with the thumb. 
12. Cut down the cast from the poured end, and fix 
the disc containing the specimen on a little 
block of wood for the jaws of the microtome, as 
directed. 
All chips of paraffin may be put back into the pot for 
remelting as often as they are cut. 70 
MEDICAL MICROSCOPY. 
VI. CUTTING SECTIONS. 
Practical microscopy has long since passed the neces- 
sity of free-hand cutting, which is now only valuable 
in history as marking the skill of some long-ago accom- 
plished workmen, the highest among them, in this as 
well as in photo-micrography, being our own lamented 
Woodward, whose free-hand sections were the wonder 
of the whole world of microscopy. 
But little progress in pathologic histology and bac- 
teriology could be made without a microtome of some 
sort, and no one wishing to engage in the examination 
of tissues can expect to do so successfully without the 
aid of this very important instrument. 
Along with the progress of more perfect microscope 
stands and objectives have come also various patterns of 
microtomes, and where all have particular claims and 
are so good, it is a difficult matter to make choice of 
the best. Indeed, it is the person who operates the 
machine that makes the good or bad performance. In 
other words, the most perfect instrument may prove a 
failure in unskilled hands, while the very poorest 
machine may make a good showing in experienced 
hands. Besides this, every worker is best acquainted 
with the instrument he uses, and, of course, can manipu- 
late it more successfully than a less experienced person. 
The best-known microtomes of American manufacture 
are probably “The Laboratory” and “Student” 
designs, manufactured by Bausch & Lomb, Rochester, 
N. Y., and the “Ryder Automatic,” manufactured by CUTTING SECTIONS. 
71 
(Cut one-third of actual size.) 
Bausch & Lomb Laboratory Microtome. 
Fig. 8. 72 
MEDICAL MICROSCOPY. 
Joseph Zentmayer, Philadelphia. These are good 
machines and, like our American microscope stands 
and objectives, fully equal to the very best of foreign 
patterns. I can manipulate either of them, I think, to 
its best capacity; but they fill very different places of 
usefulness. The Bausch & Lomb “ Laboratory Micro- 
tome ” has recently been much improved and is now 
truly first-class in every respect. The wide range of 
work for which it is fitted—cutting wet or dry, large or 
small sections—makes it almost a necessity in laboratory 
and class-room work. 
“The Ryder Automatic” is far less bulky and 
weighty, and is really a most perfect little machine. 
By using a larger and stronger knife than the common 
razor-blade attachment, its usefulness far exceeds the 
limit set by its distinguished inventor, Prof. John A. 
Ryder, of the School of Biology, University of Penn- 
sylvania, who designed it for cutting very small sec- 
tions, as a rule, for embryologic work. With this little 
machine—not occupying a space on the work-table 
larger than 4j4 x 8*4 inches, and may be lifted and 
carried with one hand—a section larger than a silver 
dollar may be cut to any desired thickness. But, as 
above stated, to reach its best capacity both as to size 
and evenness of the section, a stronger blade than a 
razor must be used. The knife manufactured by Charles 
Lentz & Sons, Philadelphia, for the Bausch & Lomb 
smaller “Laboratory Microtome,” having a cutting 
edge of 4*4 inches, exactly fills the need, and with this 
improvement the performance of “The Ryder Auto- CUTTING SECTIONS. 
73 
matic ” cannot be excelled or praised too highly, and 
is worth more than it costs. But if the cast is not a per- 
fect one, it will be impossible to make a faultless sec- 
tion, no matter what machine is employed. 
The statement on a preceding page concerning the 
short time—but four or five hours—within which a 
Fig. 9. 
Zentmayer “ Ryder Automatic Microtome.” 
permanent section-mount may be made, shows how un- 
necessary the use of the Freezing Microtome even 
in laboratory work. It is a refinement truly “ more 
ornamental than usefuland the physician who invests 
in such a machine will soon find out to his sorrow that it 
were better he had spent his money for something more 74 
MEDICAL MICROSCOPY. 
needful. I speak from experience, in having to regret 
such a purchase. 
Sharpening the knife is an important part of 
technique ; for without an edge sharp enough to “ cut 
a hair” the section will be imperfect. The slightest 
gap makes its mark in the section, hence the necessity 
of a faultless edge. Every operator should learn as 
soon as possible how to hone and strop his knife ; and 
to that end a slate hone and good strop should be a part 
of his outfit. The hone is the ordinary slate or water 
hone used by barbers. 
An important little device is the Section-flattener, 
to be fixed to the knife. By its use all imperfect sec- 
tions may be seen in the cutting, so that there need be 
no waste either of time or material. A wire with paper 
flange, devised by the author, can be fitted to any 
blade. 
VII. FIXING THE SECTION IMMEDI- 
ATELY ON THE SLIDE. 
This is a great improvement over the old method of 
dealing with the section. Instead of lifting and trans- 
ferring the section from one watch-glass and reagent to 
another, the section is at once fixed on the slide by 
means of a carbolized solution of egg albumin, made as 
follows : Beat up the white of one fresh egg in eight 
ounces of a two per cent, aqueous solution of carbolic 
acid; then filter until a clear, pearly-white solution 
results. FIXING SECTION ON SLIDE. 
A small ordinary capped bottle should be filled for 
convenient use at the work-table, and the remainder 
bottled, well-stoppered, and put away for future need. 
A camel’s hair pencil with quill long enough to be 
easily caught hold of may be kept in the capped bottle 
and used as a dropper. 
I have been using this particular fixing-medium to the 
exclusion of all others for the last six years. 
Before commencing to cut, the operator should have 
placed within convenient reach as many clean slides as 
will be needed at that sitting ; a piece of blotting paper 
containing the following diagram upon which to lay 
75 
Fig. io. 
Centering Diagram. 
the slide. Then, with the camel’s hair pencil, place a 
drop of the fixing solution at the center. The section 
is then cut, its edge caught with the fine pointed for- 
ceps, and placed on the drop of fluid at the center of 
the glass slip. A piece of clean, heavy, white filtering 
paper is laid over the section, and upon this a slip of 
smooth, oiled cardboard is pressed to squeeze out the 
excess of fluid and smoothly fix the section to the glass. 76 
MEDICAL MICROSCOPY. 
Better than oiled cardboard for such use is a slip of 
thin sheet celluloid. 
After removal of the papers, the slide is wiped and 
exposed to the air of the room to dry, say for five or 
ten minutes, after which it is placed in spirits of turpen- 
tine to get rid of the paraffin in the section. 
This method of dealing with sections—fixing them 
immediately on the slide—I have constantly employed 
for the last nine or ten years, and owe Dr. Win. M. 
Gray, of the Army Medical Museum Microscopic La- 
boratory, for the suggestion. In no other way can a 
section be safely dealt with. 
“ A method of fixing sections on the cover-glass,” 
instead of the slide, was made known to a New York 
City medical society less than two years ago by Dr. J. 
M. Byron, director of the Bacteriologic Department 
of the Loomis Laboratory, and published in Volume II 
(1892) of the Researches of the Laboratory and else- 
where ; but it is in no respect equal to the older method 
I have described. Indeed, the simplicity of the old is 
in striking and suggestive contrast with the new. 
As demonstrator of pathologic histology at the re- 
cent Pan-American Medical Congress, I had the honor 
of showing all the steps of the better method of fixing 
the section immediately on the slide. 
While by frequent whipping of the slide in turpentine 
the paraffin may be sufficiently dissolved within an hour 
to go on with the staining and finishing of the section, 
it is my habit to cut and fix the sections in the evening, 
and let the slides remain in the turpentine over night. STAINING AND FINISHING. 
77 
After removal from the turpentine the slides are thor- 
oughly washed in benzol, and wiped close to the edge 
of the section. They are then put into commercial 
alcohol “ No. 1,” and within the next five or ten 
minutes transferred to alcohol “ No. 2.” After thus 
treating a few slides, a “ No. 3 ” bottle of alcohol will 
be necessary to maintain a clear unmixed standard, in 
which the slides, with their sections duly fixed, may 
remain until it is convenient to finish them. 
The best and most beautiful nuclear pictures are made 
with borax-carmin and hematoxylin. The former is 
made as follows :— 
Best “ No. 40” carmin, I dram. 
Pulverized borax, 2 drams 
Carbolized water (2 per cent.), 4 ounces. 
Heat the solution in a glass or porcelain vessel to 
boiling, and when cool filter into a bottle with mouth 
wide enough to receive the width of a slide. 
A decolorizing mixture is necessary to bring out the 
finest effect of the stain, and may be made thus:— 
Hydrochloric acid, chem. pure, 3 drams. 
70 per cent, alcohol, ounces. 
This mixture should also be put into a wide-mouth 
bottle, for the same reason above given, namely—the 
easy handling of the slide. 
VIII. STAINING AND FINISHING. 78 
MEDICAL MICROSCOPY. 
How to Use the Stain.—With the forceps in the 
right hand, a slide is taken from the alcohol and im- 
mersed for twenty or thirty seconds—whipping it up 
and down—in the bottle of carmin. Still holding the 
slide with the forceps, it is drained for a moment on the 
mouth of the bottle and then transferred to acid alcohol 
for decolorization and proper differentiation of the sec- 
tion,—but just how long it should there remain words 
cannot tell more definitely than to say from one minute 
to a half hour, the time depending entirely upon the 
particular character of the tissue and its duration in the 
stain. Experience is the only unerring guide in the 
matter, and repeated failures, at first, should not be 
discouraging. It is generally the case that beginners 
oftener over-stain than under-stain their sections. 
The next step after use of the acid alcohol is thorough 
washing in clear water to get rid of all trace of the acid ; 
next, the slide is put back into commercial alcohol; next, 
into absolute alcohol. Then, freeing it from the forceps 
and seizing the upper left-hand corner between the left 
thumb and index finger—being careful to hold the slide 
as level as possible,—the section is then washed and 
cleared up by pouring upon it from a bottle held in the 
right hand either benzol or xylol. After the clearing 
up, the slide is cautiously wiped with a cloth all around 
the section. Then a drop of balsam and xylol let fall 
upon the center of the section, and placing upon it a 
cover-glass—either a square or circle—finishes the 
mount. 
Should it be desired to double-stain the section, the STAINING AND FINISHING. 
79 
very finest differential effect may be accomplished with 
borax-carmin, as a base, and sulphindigotate of soda, 
commonly called indigo carmin. 
After the return of the slide to commercial alcohol 
from the acid alcohol and plain water wash, a weak 
aqueous solution of sulphindigotate of soda is poured 
upon the section and allowed to remain for a half minute 
or longer; the slide is then quickly whipped three or 
four times in clean water and placed immediately into 
commercial alcohol. The subsequent steps are precisely 
those above stated in dealing with and finishing a single 
stained section. 
Solutions of sulphindigotate of soda do not keep well 
even when made with carbolized water, on which account 
but a small quantity should be made at a time. It should 
also be remembered that the color deepens when the 
section is placed in alcohol. 
Those of my readers who may wish to know some- 
thing about the refinement of treble staining are 
referred to other books for the knowledge. 
Staining with Hematoxylin.—The best formula, 
probably, is the one bearing Delafield’s name. It is 
made as follows :— 
“To ioo c.c. of a saturated solution of ammonia 
alum, a solution of one gram of hematoxylin, dissolved 
in six c.c. of absolute alcohol, is added, drop by 
drop. The solution is exposed to the air and light, in 
an unstoppered bottle, for three or four days; it is then 
filtered, and twenty-five c.c. of glycerin, and twenty- 
five c.c. of alcohol are added. The mixture is allowed 80 
MEDICAL MICROSCOPY. 
to stand until its color becomes dark, and is then again 
filtered and preserved in a bottle with a closely fitting 
stopper. It keeps well, but should not be used for two 
months after it has been prepared.” 
Recently, I have added to Delafield’s preparation, at 
the time of use—and with fine effect, I think—just 
enough sulphindigotate of soda to produce double stain- 
ing. The method is this: Take a dram or two of five 
per cent, carbolized water and add to it enough of the 
sulphindigotate of soda to give a deep bluish-green 
color ; then, to one part of this solution add seven parts 
of Delafield’s Hematoxylin. The double-stain thus 
made should be poured on the section and let stand 
from a half hour to an hour and a half, or longer if ne- 
cessary, to give sufficient color. After staining, the 
slide holding the section is immersed in a beaker of weak 
acidulated water—made by adding a few drops of nitric 
acid—and washed until sufficient decoloration has taken 
place. It should remain in the acidulated water until 
the section shows a clear, deep, sky-blue color ; then it 
should be thoroughly washed in clear water, and after- 
ward transferred to commercial alcohol, to remain at 
least twenty minutes to deepen the color. From the al- 
cohol it is to be treated and finished in the same man- 
ner as for a simple carmin mount. 
This hematoxylin double-stain differentiates the cell- 
inclusions— “chromatin,” “protozoon parasites,” 
“coccidia,” “ psorosperms,” “ Rhopalocephalus car- 
cinomatosus,” or whatever else called—in cancer-cells 
in a surprising manner, giving better result, I think, FORMULARY. 
81 
than the Ehrlich-Biondi method, or any other with 
which I am acquainted. 
With these two stains—borax-carmin and Delafield’s 
hematoxylin with sulphindigotate of soda—may be 
demonstrated, in ordinary tissue-work, the very finest 
histologic details, and sufficient for all practical pur- 
poses, both in the hands of experts and beginners. Be- 
sides them, I could easily give enough formulae to fill a 
score of pages, yet accomplish nothing more than to 
utterly confuse the beginner, who, to be profited in the 
highest degree, should be led by the simplest methods 
to the fullness of technique. 
IX. FORMULARY. 
The Anilin Dyes.—Ehrlich first drew attention to 
the fact that these colors are divisible into two main 
groups: One, in which the staining principle is an acid; 
the other, basic—such as methylene-blue, fuchsin, gen- 
tian violet, methyl-violet, vesuvin, etc. These im- 
portant stains, and many other wonderful products, are 
coal-tar derivatives, serving the science of bacteriology 
and every day aiding its progress. 
With either an alcoholic or aqueous solution of these 
dyes all varieties of bacteria may be more or less deeply 
stained. The basic anilin stains differentiate very 
sharply both the cell nuclei and the bacteria, while most 
of the acid anilin stains are simply “ nuclei stains,” and 
worthless in differentiating bacteria. 
Gentian Violet is a very intense stain, and in its 82 
MEDICAL MICROSCOPY. 
use there is more danger of over staining than under 
staining. 
Fuchsinis also an intense stain, pleasant to the eye, 
and not so apt to overstain as gentian violet. 
Methyl-violet colors less intensely than either of 
the above named stains, also is less durable. 
Methyl-blue is the feeblest of the four colors above 
listed, and therefore requires longer time to produce 
thorough staining; but it has a wide range of usefulness 
and is very durable. 
Bismarck Brown, orVesuvin, has a narrow field 
of usefulness, being mostly used as a contrast stain with 
gentian violet. 
These five basic anilin colors fill the most important 
place in bacteriologic demonstrations, and are sufficient 
for all practical purposes. The technique for the em- 
ployment of one, with but few exceptions, will stand for 
all, both in cover-glass preparations and section mounts. 
Many valuable combinations and methods have been 
given and indorsed by distinguished experts in mi- 
croscopy and bacteriology; but as it is not within the 
scope of this volume to reproduce them all, I shall limit 
the description to those only which I am in the habit of 
using, referring the reader to the various text-books 
for all the knowledge on the subject. 
No. 1.—Carbolized Solution Methyl-blue.— 
Ten c.c. of absolute alcohol are poured over 1.5 grm. of methy- 
lene-blue in a mortar and 100 c.c. of a five per cent, aqueous solution 
of carbolic acid then added, and the whole rubbed together until 
the blue is entirely dissolved. FORMULARY. 
83 
No. 2.—Aqueous Solution Methyl-blue.— 
Methylene-blue, 2 grm. 
Alcohol, 15 c.c. 
Water, 85 c.c. 
No. j>.—Koch’s Solution Methyl-blue.— 
Concentrated alcoholic solution methyl-blue, 1 grm. 
Ten per cent, potash solution,* 2 c.c. 
Distilled water, 200 c.c. 
No. 4.—Loffler’s Solution Methyl-blue.— 
Concentrated alcoholic solution, 30 c.c. 
Potash solution, 1-10,000 100 c.c. 
No. 3.—Chenzynski’s Solution.— 
Concentrated aqueous solution methyl-blue, 40 grm. 
1-2 per cent, solution of eosin in 70 per 
cent, alcohol, 20 grm. 
Water, 40 “ 
12 drops of a 20 per cent, solution potash, aqueous. 
No. 6.—Plehn’s Solution.— 
Concentrated aqueous solution methyl-blue, 60 parts. 
1-2 per cent, solution eosin in 75 per cent. 
alcohol, 20 “ 
Distilled water, 40 “ 
12 drops of a 20 per cent, solution caustic 
potash. 
No. 7.—Unna’s Borax Methyl-blue.— 
Borax, 1 part. 
Methylene-blue, 1 “ 
Water, 100 parts. . 
For tubercle and lepra bacilli, stain sections for five minutes; then 
* Potash solution, 1-10,000. 84 
MEDICAL MICROSCOPY. 
into five per cent, aqueous solution pot. iod. with a crystal of iodin 
added; then rinse in alcohol until blue cloud forms; then differ- 
entiate in creosote. 
No. 8.— Unna’s Stain for Microorganisms in the Skin.— 
Sections to be first stained in carmin, then for two minutes in 
borax methyl-blue, rinsed in water, for a few seconds in aqueous 
solution arsenic acid, next in alcohol, next in xylol, then balsam. 
No. g.—Noniewicz-Loffler-Unna Method for Glanders 
Bacilli.— 
Sections are transferred from alcohol to concentrated aqueous so- 
lution methyl-blue for two to five minutes, then rinsed in water, then 
decolorized in a mixture of 75 parts of 1-2 per cent, acetic acid and 
25 parts of 1-2 per cent, aqueous solution of tropaeolin, washed in 
water, dehydrated, cleared with xylol. 
No. 10.—Anilin Oil Methyl-blue.— 
Methylene-blue, 1 grm. 
Anilin oil, 10 c.c. 
Gentian Violet.— 
No. ii.—Ehrlich’s Solution.— 
Concentrated alcoholic solution gentian vio- 
let 5 c.c. 
Anilin water, ioo c.c. 
No. 12.—Aqueous Solution.— 
Gentian violet, 2.25 grm. 
Distilled water 100 “ 
Fuchsin (rosanilin hydrochlorate). 
No. ij.—Carbolized Fuchsin (Nielsen's). 
Fuchsin, i grm. 
Alcohol, io c.c. 
5 per cent, carbolized water, ioo c.c. FORMULARY. 
85 
No. 14.—Aqueous Solution.— 
Fuchsin (rosanilin hydrochlorate), .... 2 grm. 
Alcohol, 15 c.c. 
Water, 85 c.c. 
No. 13.—Gibbes’ Double Stain.— 
Rosanilin hydrochlorate, 2 grm. 
Methylene-blue, 1 grm. 
Triturate in a glass mortar; then dissolve 
anilin oil, 3 c.c. 
In absolute alcohol, 15 c.c. 
Add this to the stains, then slowly add dis- 
tilled water, 15 c.c. 
No. 16.—Carbolized Double Stain.— (Reeves.) 
Rosanilin hydrochlorate, 2.5 grm. 
Methylene-blue, 1.5 grm. 
Absolute alcohol, 20 c.c. 
Anilin oil, 3 c.c. 
Shake well and add slowly 5 per cent, car- 
bolized water, 30 c.c. 
Methyl-violet. 
No. iy.—Koch’s Solution.— 
Concentrated alcoholic solution, 11 c.c. 
Absolute alcohol, 10 c.c. 
Anilin water, ... »oo c c. 
No. 18.—Methyl-violet, 2.25 grin. 
Carbolized water, 100 c.c. 
Bismarck Brown. 
No. ig.—Concentrated Solution.—Equal parts of glycerin 
and carbolized water. 86 
MEDICAL MICROSCOPY. 
No. 20.—Aqueous Solution.— 
Bismarck brown, 2 grm. 
Alcohol, 15 c.c. 
Carbolized water, 85 c.c. 
No. 21.—Borax Carmin. (See page 77.) 
No, 22.—Hematoxylin Double Stain. (See page 80). 
No. 23.—Weigert’s Fibrin Stain. 
The section, hardened in alcohol, is stained for an hour in satur- 
ated anilin gentian violet (No. 11), rinsed in 0.7 per cent, solution 
of table salt. After washing in water and partially drying the sec- 
tion by means of filter paper, iodin water (No. 25) is dropped upon 
it and thus flooded for a minute or two, after which filter paper is 
again applied to absorb the superfluous fluid, and anilin oil poured 
on for the purpose of decolorizing and anhydrating it. The anilin 
oil must be replaced two or three times, or as long as it becomes dark, 
by fresh oil, and then entirely replaced with xylol, after which comes 
balsam and the cover-glass. 
ATo. 24.—The Author’s Method.— 
Concentrated alcoholic solution of safranin, . 
Concentrated aqueous solution of safranin, . 
equal parts. 
The section should be flooded on the slide with this solution for 
a half hour, after which it is washed for a few minutes in commer- 
cial alcohol. A weak solution of sulphindigotate of soda in 5 per 
cent, carbolized water is then poured on and allowed to remain a 
minute or two, or until the fibrin threads are sufficiently colored. 
After the use of the sulphindigotate of soda, the section is whipped 
several times in clean water, and then transferred to alcohol, and 
cleared with benzol or xylol. By this method a pretty orange- 
red color is given the nuclei, while the fibrin threads are stained 
more or less deeply a bluish-green. FORMULARY. 
87 
In preparing the safranin stain, great care should be exercised to 
have a reliable dye—Griibler’s, if possible. 
As a nuclear stain safranin, in the above proportions, may be em- 
ployed in studying the cell-structure of morbid growths, the process 
of karyokinesis, and also epithelial structures, but it cannot take the 
place of hematoxylin (No. 22) for the same uses. 
No. 23.—Gram’s Solution.— 
Iodin 1 part. 
Iod-pot, 2 parts. 
Distilled water, 300 parts. 
No. 26.—Gram’s Method Modified.— 
First of all, stain the section safranin (No. 24) for a few minutes; 
next, rinse in water; next, stain with methyl-violet (No. 17 or 18) 
for five minutes; next, wash in water; next, Gram’s solution for a 
minute and a half; next, pour off the solution and dry the surface of 
the section with filter paper; next, anilin oil in sufficient changes to 
decolorize and anhydrate the section; next, thorough washing in 
xylol to get rid of all trace of the oil; and then balsam and the 
cover-glass. 
No. 27.—Anilin Oil Solutions.— 
Anilin Oil Solutions of methyl-blue, safranin, auramin, acid 
violet, etc. 
These solutions may be employed either for restoring too much 
waste in decolorizing, or as second stains. 
No. 28.—Anilin Water.— 
Anilin oil, 12 drops. 
Distilled water, 6 c.c. 
Shake well together and pass through a moist filter. 
No. 2g.—Flemming’s Solution.— 
To four parts of a two per cent, aqueous solution of osmic acid, 
add 15 parts of one per cent, solution of chromic acid, and one part 
of glacial acetic acid. 88 
MEDICAL MICROSCOPY. 
After remaining in this mixture for two or three days, the tissue- 
specimen should be washed in water for three to six hours, and 
finally hardened by successive immersions for 24 hours each in 30, 
60, and 96 per cent, alcohol. 
No. 30.—The Ehrlich-Biondi Triple Stain.— 
Dr. Sims Woodhead prepares the stain as follows :— 
Saturated solution of methyl-green, .... 5 c.c. 
“ “ “ methyl-orange, ... 10 c.c. 
“ “ “ acid fuchsin, .... 2 c.c. 
To each of the above substances add about 40 volumes of water, 
to avoid the formation of a precipitate; then mix all together. 
Stain the section for about 15 minutes to 12 hours, rinse in 1 
per 1.000 acetic acid, wash in dilute and then for one minute in 
absolute alcohol, and finally immerse in xylol or benzol, and mount 
in xylol-balsam. 
No. 31.—Malachowski’s Method.— 
1-2 grm. each of borax and methyl-blue in 100 c.c. of water. 
No. 32.—Toison’s Staining Fluid.— 
Distilled water, 160 c.c. 
Glycerin, 30 c.c. 
Sulphate soda, 8 grm. 
Chlorid sodium, I grm. 
Methyl-violet, 0.025 grm- 89 
BACTERIOLOGY. 
X. BACTERIOLOGY. 
The germ theory of disease, now so progressive and 
popular, is not a modern creation. It is nearly two 
and a half centuries old; indeed, almost coeval with 
the discovery of the microscope, whose astonishing 
revelations of bacterial life swarming in stagnant waters, 
stale infusions of vegetable and animal substances, also 
in the secretions from the mouth and accumulations 
about the teeth and gums, gave origin and currency 
long, long ago, to the belief that definite microorgan- 
isms were active factors of many contagious diseases. 
The proof that a given organism is the cause of a par- 
ticular disease can only be shown as follows:— 
First. “ An organism of a definite form and with 
definite characteristics of life-history must always be 
found in the blood or in the affected parts of the animal 
body. 
Second. “The blood or the affected parts contain- 
ing these organisms, when inoculated into another 
animal of the same species, must produce the same 
disease. 
Third. “Treatment of the blood or affected parts in 
such a manner as to destroy the microorganisms present 
in them must also destroy their power of causing disease 
in another animal. 
Fourth. “ When the diseased parts are inoculated on 
suitable soil outside the animal the microorganisms 
grow, and can be indefinitely propagated on similar 
soil. 90 
MEDICAL MICROSCOPY. 
Fifth. “When in this manner the organisms have 
been separated from the remains of the animal sub- 
stances in which they were embedded, their inoculation 
on a suitable animal must again produce the disease, 
the same organisms being also found in the diseased 
parts.”—Cheyne. 
In numerous well-known diseases this sort of severe 
experimental proof has been furnished, anthrax being 
the highest type of absolute proof. 
In another class whose infectious character and clini- 
cal history are well understood, the invading specific 
microorganism has not yet been isolated,—yellow fever, 
for example. 
Again, the cultivation of some pathogenic organisms 
has not yet been successful—relapsing fever, syphilis, 
and Plasmodium malarice. 
It would far exceed both the purpose and limits of 
this book to attempt even a hurried run over the no- 
menclature and purely theoretical study of the various 
classifications that have been proposed in recent years. 
Medical men generally are only interested in the sub- 
ject of bacteriology from a purely etiologic standpoint, 
leaving all else to those who have inclination and time 
to spend in its search. 
Bacteria—Schizomyceles, or fission-fungi are the 
lowest members of the vegetable kingdom, closely re- 
lated to the lower A/gce. They are colorless cells of a 
transparent or glassy appearance, with enveloping mem- 
brane having protoplasmic contents but no nucleus. 
The presence or absence of the cell-capsule or pellicle BACTERIOLOGY. 
91 
sometimes serves to distinguish between microorganisms 
which strongly resemble each other, as in the case of the 
comma bacillus of Asiatic cholera, which forms a pellicle, 
whereas in Cholera nostras the Finkler-Prior bacilli do 
not. Interiorly, the cell has usually a homogeneous 
appearance, but sometimes contains oil-like granules. 
The bacteria divide themselves in a series of species, 
well defined by growth and form, which do not run into 
each other. Of the various forms in which they appear 
we know the globular bacteria, or Micrococci—found 
either singly or united in groups. When single, they 
are called Monococci; if congregated in masses, Staphy- 
lococci ; if joined in pairs and fours, they are distin- 
guished respectively, Diplococci, double, and Tetracocci, 
four. If united in blocks of eight, they are named 
Sarcina ; if linked together and chain-like, the name is 
Streptococci. 
Bacilli, or rod-shaped bacteria, are of various sizes 
and lengths, the smallest yet discovered being the in- 
fluenza bacillus. Their ends are variously shaped, 
sometimes sharply cut, in other instances rounded off, 
and more or less pointed. The bodies of the rods also 
differ in thickness as well as in lengths, sometimes larger 
at the middle than at the ends, or they may have the 
drumstick shape, as seen in the tetanus bacillus. 
Spirilla, or screw-like bacteria, are subdivided into 
comma-bacilli or Vibrios and Spirochetce, the latter 
mainly distinguished by their thread-like form and 
flexibility ; the former, by its strings of cells resembling 
spirilla. (Schenk.) 92 
MEDICAL MICROSCOPY. 
Fig. ii. 
Sarcina (Pocket-cocci). 
■ Cocci. 
Staphylococci. 
Diplococci. 
Streptococci. 
Tetracocci. 
Zooglea. 
Ciliated cell. — 
Slender bacilli. 
Short “ 
' Diplococcus. 
Spider cell. 
Bacilli in chains. 
With 
capsules 
Tetracoccus 
Vibrios (spirilla). 
Monococci 
Comma bacilli. 
Centrally situated _ 
spores. 
Spirochete. 
Knobbed bacteria with 
terminal spores. 
Clostridia forms. = 
Forms of Bacteria. 
(From Schenk, magnified about 700 times.) Many of the bacteria possess the power of movement 
and are more or less active. This, however, must not 
be confounded with oscillatory movement nor molecular 
(Brownian) movement. True movement is accomplished 
by means of flagella with which the microorganism is 
provided. These may spring from either end or both 
ends of the rodlet. 
The name Schizomycetes, or fission-fungi, is expressive 
of the mode of multiplication of bacteria. When the 
individual microorganism has reached maturity, a 
sharp line makes its appearance at the center and 
forms the division ; after that the two parts free them- 
selves from each other and become independent organ- 
isms. In that way the multiplication goes on from 
beginning to end. Upon the play of the daughter- 
cells depends the character of the grouping. If they do 
not become disjointed, the cells remain connected 
either in strings, clusters, or chains. 
Another mode of propagation is by the development 
of spores, which have remarkable power of resisting the 
influence of either extremes of temperature or the action 
of chemicals. So permanent and important is this mode, 
that it has been proposed as a basis for a scientific ar- 
rangement of the bacteria. Take, for example, the 
anthrax bacillus. While the rodlet is comparatively a 
tender organization which quickly perishes at a tem- 
perature above 6o° C., the spore is able to resist all 
the forces nature can marshal against it, either chemical 
or physical, save the one exception of sunlight and the 
actual flame. 
BACTERIOLOGY. 
93 94 
MEDICAL MICROSCOPY. 
Spores, in most cases, occupy the center of the 
rodlets, but in a few varieties they occupy the end. 
This is well seen in the tetanus bacillus. When 
occupying the center, the bulging caused by its pres- 
ence may give the organism a spindle-shape, or the 
clostridium form. 
Again, bacteria may be classed and distinguished 
according to their respective capability of living and 
multiplying upon dead substances or upon living matter 
—saprophytes and parasites. The parasites have been 
subdivided into obligate and facultative. The former 
are true parasites, thriving and multiplying only in the 
living body, and quickly perish when apart from living 
material. Facultative parasites can adapt themselves to 
either condition—live and multiply either within or 
without the living body. In other words, they are 
ambigenous. 
Besides the above-named distinctions of class and 
varieties, there are aerobes and anaerobes. The former 
cannot grow except in the presence of oxygen, while 
those microorganisms whose multiplication occurs only 
in the absence of oxygen are denominated obligate 
anaerobes. When growth may occur either in the 
presence or absence of oxygen, the microorganism is 
said to be a facultative anaerobe. 
The products of metabolism are also properties of 
bacteria. A large number of bacteria generate color- 
ing matter while they are themselves colorless and trans- 
parent ; the pigment being merely the formed product 
of their metabolism under the influence of light. Again, many species throw off odorous products, and some of 
the anaerobic class generate and exhale very offensive 
putrefactive gases, ammonia, sulphureted hydrogen, 
etc. 
Again, besides peptonizing albuminoid substances, 
some microorganisms possess the faculty of resolving 
organic bodies into their simplest elements, some break- 
ing up the chemical combination of albuminoid bodies, 
causing putrefaction ; others, inducing fermentation ; 
while a few others, in consequence of molecular activ- 
ity of their protoplasm, are phosphorescent, or have 
the property of becoming luminous in the dark. 
Up to the present time, 489 different microorgan- 
isms have been isolated and variously classified. The 
general knowledge on the subject, derived from Stern- 
berg’s “Manual of Bacteriology,” may be epitomized 
as follows :— 
BACTERIOLOGY. 
95 
Pathogenic micrococci, 44 
Non-pathogenic micrococci, 78 
Pathogenic bacilli, 108 
Non-pathogenic bacilli, 174 
Pathogenic spirilla, 6 
Non-pathogenic spirilla, 21 
Leptotrichea and cladotrichea, 8 
Not classified, 50 
Asiatic Cholera.—Standing at the head of the 
class of pathogenic bacteria is that most dreadful of all 
disease-producing parasites, the comma bacillus of Asi- 
atic cholera (Koch). A definite and morphologically 
distinctive microorganism found in the alvine dis- 96 
MEDICAL MICROSCOPY. 
charges and intestinal walls, rarely in the vomit, never 
in the breath, urine, tears,' or other secretions, nor in the 
blood; body from 0.8 to 2.0 long, or about half as 
long as the tubercle bacillus, sometimes curved and so 
connected at the ends as to form S shapes and half cir- 
cles, or running out into long or short spirals f resem- 
bling the spirilla of relapsing fever, motile and armed 
with a single flagellum, by which it whirls through the 
microscopic field “ like a swarm of dancing gnats,”— 
aerobic and grows rapidly in gelatin and blood serum 
at a temperature of 30° or 40° C.—multiplies by divi- 
sion—not yet known whether it is spore-producing or 
not—acids unfavorable to its growth,—easily killed by 
drying: thermal death-point 520 C. 
Methods of Artificial Culture.—Koch’s method 
of plate-cultivation with nutrient gelatin has been gen- 
erally accepted as the best. Thus cultivated, the colonies 
commence to form in 24 hours, or less. They make 
their appearance in small, white specks or dots, sinking 
below the surface, resembling so many air-bubbles, and 
may be seen with the naked eye. When magnified 100 
diameters, the field appears as if strewn with bits of 
glass. As the culture grows older the gelatin becomes 
more and more thoroughly peptonized, and during the 
liquefying process a disagreeable odor is emitted. 
* fi the micron or micromillimeter = .001 mm., of an 
inch, nearly. 
f Strict systematicians look upon the “bacillus ” as only a frag- 
ment of a real Spirillum. BACTERIOLOGY. 
97 
The nutrient gelatin may be prepared as follows:— 
Take one pound of beef, free as possible from fat, 
and chop it up finely; then transfer the mass to a glass 
or porcelain vessel and add a quart of distilled water. 
Shake well the mixture and place it in a refrigerator for 
24 hours. After thus standing, it is again well shaken 
up and the liquid portion filtered by squeezing through 
a linen cloth. Then to the reddish-colored filtrate add 
enough distilled water to make a quart mixture, and im- 
mediately put into it 
5 grms. of common salt, 
lo grms. of dry peptone, 
too grms. of best white edible gelatin. 
Shake up well, and place the vessel in a water-bath 
with gentle heat for a half hour, or until the gelatin is 
completely dissolved. 
The test should now be made to make sure of exact 
neutrality of reaction, and, if necessary, sodium car- 
bonate added to produce such result. The liquor is 
then heated for an hour in the water bath and tested 
again for its reaction. In about ten minutes before the 
boiling is completed, the white of an egg, with the 
shell, is added, and the mixture then filtered while hot, 
through paper and a funnel specially fitted for the pur- 
pose. Should the filtrate show the least turbidity, it 
must be put back into the heating vessel and boiled a 
second time for at least ten minutes, then refiltered. 
But it should be remembered that too long boiling pre- 
vents the gelatin from solidifying. 98 
MEDICAL MICROSCOPY. 
This formula may be modified at will by the addition 
of organic or inorganic substances, and will serve for 
the cultivation of all pathogenic bacteria which grow at 
the ordinary room temperature. 
How to Make the Plate-culture.—A glass plate, 
or one of iron, having first been carefully sterilized, is 
cooled and then exactly leveled on a tripod constructed 
for that special purpose. Upon this plate is laid 
another sterilized plate, either of glass or iron, but 
Fig. 12. 
Koch’s Plating Apparatus as arranged for “ Setting ” Agar or Gelatin Plates 
smaller than the bed-plate, and upon it the nutrient 
gelatin is poured, as evenly as possible, and at once 
covered with a sterilized bell-glass. 
After the gelatin coating has sufficiently cooled and 
set to permit movement, the plate is transferred to a 
glass dish in which it is placed on sterilized blotting 
paper wet with corrosive sublimate solution, and covered 
with a sterilized bell-glass. 
The inoculations are then made with a platinum BACTERIOLOGY. 
99 
needle, also duly sterilized and charged with a particle 
of the infective material, with which the gelatin surface 
is pricked here and there. 
Agar-agar.—In all cultures which do not require 
a lower temperature than blood-heat, this medium may 
be used. It is prepared in the same way as nutrient 
gelatin, only that 1.5-2 per cent, of finely divided agar- 
agar is used. 
Nutrient Potato.—Sterilized potatoes make a con- 
venient culture-garden, and are of great service in study- 
Fig. 13. 
Oese or Platinum Needles, Straight, Hooked, Looped. 
ing pathogenic bacteria. The bacterium of cholera, 
glanders, enteric or typhoid fever, diphtheria, etc., 
develops on potato, and in some cases in a highly char- 
acteristic manner, particularly enteric or typhoid fever. 
When this nutrient substance is employed, potatoes with 
smooth skins and but few eyes should be selected. They 
are at first scrubbed and freed from sand or dirt of any 
kind with a brush, and afterward placed for an hour in a 100 
MEDICAL MICROSCOPY. 
five per cent, corrosive sublimate solution, and then 
boiled for a half hour. They are next cooled on a 
sterilized plate, cut into halves with a knife that has also 
been properly sterilized, and otherwise made ready for 
inoculation. For further instructions in the technique 
of culture media the reader is referred to any standard 
text-book on bacteriology, particularly Dr. Sternberg’s 
great work. 
Cover-glass Demonstration.—The cholera mi- 
croorganism is not always an easy matter to demon- 
strate in cover-glass preparation from the typical rice- 
Fig. 14. 
Damp Chamber for Plate Cultivations. 
water stools, owing to the “garden full of weeds” 
which is usually present in the microscopic field ; but 
an accidental good catch may now and then be made, 
and is therefore worth trial in all cases. Where this 
fails, the method of plate-cultivation above particularly 
described should be promptly resorted to, for upon the 
correct diagnosis of the “suspicious case ” may depend 
the safety of a whole community. BACTERIOLOGY. 
101 
A speck of the heaviest flaky matter is picked out, with 
a large needle or other instrument, from the stool and 
smeared on a clean cover-glass. Upon this is then laid 
another like clean cover and both pressed together in 
order to give an even coating to both glasses. The 
squeezed-out matter around the edges is then wiped off, 
and the two pieces carefully slipped apart. After drying 
in the air of the room for a few minutes, they are caught 
with the forceps and whipped several times—the smeared 
Fig. 15. 
Comma Bacillus of Asiatic Cholera. 
side up—through the flame of a spirit lamp to fix the 
serum-albumin and microorganisms in the film. After 
this preparation of the covers, they are laid on blotting 
paper, coated side up, and covered with a bell-glass to 
wait for the application of the stain, to be used as fol- 
lows : Put into a small test-tube, having a lip—and fixed 
in a holder—a little alcoholic solution of fuchsin, methy- 
lene-blue, or violet, and double its bulk with five per cent, 
carbolized water. Shake the mixture, and hold the tube 102 
MEDICAL MICROSCOPY. 
over the flame of a spirit lamp until the fluid begins to 
steam, then lift the bell-glass, pour on each of the glasses 
just enough of the stain to fully cover their surfaces, and 
replace the bell-glass. 
After standing thus flooded with the stain for one or 
two minutes, catch the edge of the cover-glass with the 
forceps and drain off the waste staining fluid by tilting 
the glass on a blotting paper ; then, while still holding 
Fig. 16. 
Finkler-Prior Bacillus of Cholera Nostras. 
the glass with the forceps, wash thoroughly in water, 
and stand it on its edge on blotting paper to drain and 
dry. Before mounting on the slide, the covers should 
be thoroughly dried by heat to make sure they are in con- 
dition to receive the drop of balsam and xylol to clear 
up the film and seal them to the slide. 
After finishing the mount, if the specific microorgan- 
ism is present, it may be discovered with a Bausch & 
Lomb i—12 oil immersion, aided by an Abbe condenser. BACTERIOLOGY. 
103 
Bacillus of Cholera Nostras.—The Finkler-Prior 
bacillus is larger and thicker than the Comma bacillus of 
Koch, and, because of its resemblance to the micro- 
organism of true Asiatic cholera, may be mistaken for 
it in a cover-glass preparation from the stools. In case 
of doubt, the culture-method should be resorted to, by 
which the question can soon be settled, namely, whether 
it is the Vibrio proteus of Cholera nostras, or the Comma 
bacillus of Asiatic cholera, which is present. The cul- 
ture on a nutrient gelatin plate liquefies so rapidly and 
extensively after inoculation with the Vibrio proteus that 
a mistake should not be made in interpreting the correct 
result of the culture. Even on potato, the crucial test 
may be made. On this culture-medium, the Vibrio 
proteus grows at ordinary temperatures, while in genuine 
or Asiatic cholera the Comma bacillus only grows at 
incubator temperature. Thermal death-point 50° C., 
same as Asiatic cholera. 
Enteric or Typhoid Fever.—At present the con- 
sensus of opinion among bacteriologists accepts without 
hesitation Eberth’s bacillus, discovered in 1880, as the 
true pathogenic factor of enteric or typhoid fever. The 
etiologic relation of the microorganism has been 
abundantly confirmed by the researches of Klebs and 
Eppinger, Koch, Friedlander, Pfeiffer, Frankel and 
Simmonds, Meyer, Gafifky, Sternberg, and other dis- 
tinguished authorities in bacteriologic science the 
world over. Sternberg says : “ No other organism has 
been found, after the most careful search, in the deeper 
portions of the intestinal glands involved in this disease, 104 
MEDICAL MICROSCOPY. 
or in the internal organs; on the other hand, this bacil- 
lus has been demonstrated to be constantly present. It 
is undoubtedly present during the life-time of the 
patients, and is found in greater abundance in those 
cases which terminate fatally at an early date. It is not 
a putrefactive organism, and is not developed in the tis- 
sues post-mortem, although it has been shown by Frankel 
and Simmonds that it multiplies rapidly in the spleen 
after death, up to the time that putrefactive decomposi- 
tion commences.” 
While the spleen is the organ in which the bacillus 
Fig. 17. 
Bacillus of Typhoid Fever.—Rods and Spider-Cells 
has been more frequently found, it also invades the 
liver, kidneys, and mesenteric glands. Riitimeyer and 
Newhauss have stated that it may also be found in the 
blood of such typhoid patients, taken from the rose- 
spots, and that they succeeded in cultivating the 
specific microorganism from it. Further confirmation 
on this point, however, seems necessary, as the more 
recent researches of Janowsky show that the bacillus is 
very rarely present in the blood during life, and its de- 
tection should therefore not have much weight in the 
resources of the differential diagnosis. It is always extremely difficult to demonstrate its 
presence in the feces; hence the necessity of appeal to 
artificial cultivations. The morphologic description 
given by various authors is uniform in every essential 
particular,—namely: that the microorganism is “three 
times as long as broad, with rounded ends, may grow 
to long threads as well as be found in short rods, motile 
and probably possesses flagella, and resists the action 
of anilin colors to a greater degree than most similar 
organisms.” The high degree of motility is well ac- 
counted for when Loffler’s “spider-cells” are placed 
under a power of 1.200. With such amplification the 
rodlets present really the appearance of spiders. 
Of all the diagnostic characters given, says Sternberg, 
“ the peculiar growth upon potato, first pointed out by 
Gaffky, is the most important. This serves to distin- 
guish the typhoid bacillus from similar organisms which 
in their morphology and growth upon gelatin or upon 
agar-agar closely resemble it. The rods, as first pointed 
out by Eberth, have rounded ends, but this is by no 
means peculiar to the typhoid bacillus. They vary 
greatly in length, and while the prevailing form is that 
of a short rod three or four times as long as broad, fila- 
ments of considerable length are developed in cultures 
made in gelatin or upon potato.” 
When a positive decision is required concerning the 
presence or absence of the microorganism, the potato 
culture-test may be employed, but it cannot be relied 
upon alone. In three or four days, at room temperature, 
and provided the slices of potato have an acid reaction, 
BACTERIOLOGY. 
105 106 
MEDICAL MICROSCOPY. 
—for which the culture should always be previously 
tested,—there appears on the surface a moist, pearly- 
looking gloss which is peculiar to the growth of the 
typhoid bacillus on potato. In case of doubt, a further 
means of recognition has been proposed by Chante- 
messe and Widal, namely, the carbolized nutrient gela- 
tin-test. These authorities are responsible for the 
statement that it is a peculiarity of typhoid fever bacilli 
that they thrive on a nutrient gelatin which has been 
mixed with 2 per 1.000 of carbolic acid, whereas all 
other microorganisms perish on this mass. It thrives 
whether oxygen is excluded or not, but is more luxuriant 
in the presence of this gas. 
According to Gaffky and Birch-Hirschfield, spores 
occupying the ends of the rodlets are formed in the 
course of three or four days at a temperature of 33°-4o° 
C., and at a temperature lower than 20° C. sporulation 
ceases. This statement has been antagonized by Buch- 
ner, Janowsky, and Sternberg, who believe that the 
refracting granules described by Gaffky as spores are 
not reproductive bodies—and for this reason they are 
utterly destroyed by a temperature of 6o° C., while 
spores are distinguished by their resistance to heat and 
other destructive agencies. In fact, according to Buch- 
ner, the bacilli containing these refractive granules are 
even less resistant than fresh cultures in which they are 
not present; and he looks upon them as representing a 
degeneration of the protoplasm of the cells. (Stern- 
berg.) 
The thermal death-point of this bacillus is 56° C. BACTERIOLOGY. 
107 
Cover-glass preparations may be made as directed for 
the demonstration of the cholera bacillus, except that 
the time of staining should be lengthened from ten to 
fifteen minutes. 
Staining Sections.—It has already been stated that 
the bacillus of enteric or typhoid fever is refractory to 
the anilin stains, as ordinarily used. By a very simple 
method I have been able to overcome completely such 
difficulty in staining, and by it produced the most sat- 
isfactory results—not only in demonstrating the typhoid 
bacillus, but as well other specific microorganisms in tis- 
sue. In fact, by the method now to be described (for 
the first time, I believe) there is not the least danger— 
I should have said possibility—of washing out too much 
of the stain in the process of dehydrating the section. 
It completely bridges over “the weak point,” stated 
by Dr. Kiihne, “ in the processes hitherto used, viz., the 
dehydration of already differentiated sections in alco- 
hol, by which under certain circumstances the decolor- 
izing properties of the fluid must have had a very bad 
effect. “ I tried,” says Dr. Kiihne, “to minimize this 
undesirable accessory effect of the alcohol by adding to 
it some of the stain in which the sections had been pre- 
viously stained, and by these means a part at least of the 
staining matter removed by the alcohol was replaced. 
Afterward anilin oil was recommended by Weigert as a 
means of dehydration,” and to meet the emergency. 
The “simple method” with benzol, referred to 
above, is so successful that indeed both absolute alcohol 
and anilin oil may be dispensed with as dehydrating 108 
MEDICAL MICROSCOPY. 
agents in that class of fission fungi fatally faded by al- 
cohol. By its use deep staining is unnecessary. 
The material having been properly hardened in alco- 
hol, cast in paraffin, the section cut and immediately 
fixed on the slide with a carbolized solution of egg- 
albumin, then freed of all trace of the embedding mate- 
rial by soaking in spirits of turpentine, next thoroughly 
washed in benzol followed with alcohol,—in other 
words, the section having been made ready for staining, 
—the procedure is as follows: Take a small quantity of 
“ Loffler’s Solution,” or, instead, equal parts of a strong 
alcoholic solution of methyl-blue and five per cent, car- 
bolized water, in a test-tube in the manner described 
on a preceding page, and steam it over the flame of a 
spirit lamp. Then, taking a slide holding the section 
from the beaker or bottle of alcohol, the under surface 
and ends are wiped quickly with a linen handkerchief, 
after which it is placed section side up on a piece of 
blotting paper. The hot staining fluid is then poured 
upon the section, and the slide covered with a bell- 
glass. 
The section should remain covered with the stain for 
ten or fifteen minutes. Then, catching the slide with 
the forceps and draining off the surplus stain, it is 
washed in distilled water for a few seconds and next 
wiped with a cloth, especially the under side. A bit or 
square of fine, thick, white filter-paper is then pressed 
upon the section to absorb the water and stain on its 
surface. 
The section is now flooded with benzol—the upper BACTERIOLOGY. 
109 
left-hand corner of the slide being held between the 
thumb and index finger of the left hand—for a minute 
or so; next, the benzol is poured off and another piece 
of the filter paper applied ; then flooded again with ben- 
zol for half a minute, after which it is replaced with 
xylol. This process of dehydration should be con- 
tinued until the section is well cleared up, after which 
a drop of balsam and xylol is placed upon the section 
and the cover-glass laid on. 
It may be here remarked that benzol not only has 
the capacity to replace spirits of turpentine, but posses- 
ses great affinity for water, on which account it is of 
priceless value in the demonstration of some particular 
forms of bacteria in tissue. 
By this method, absolute alcohol is not needed in 
dehydrating sections, and thus treated microorganisms 
that do not stain well by the old methods, or are easily 
faded with alcohol, hold fast all the color necessary to 
make their demonstration complete. 
Pulmonary Tuberculosis.—Happily, there no 
longer exists any excuse for uncertainty concerning the 
true condition in a case of pulmonary tuberculosis. 
Koch’s discovery of the tubercle bacillus has made possi- 
ble an unerring diagnosis, even in the incipient stage 
and before there is manifested a single physical sign to 
give warning of the presence of the disease. Recogni- 
tion of this inestimable truth should quicken the physi- 
cian’s sense of his great responsibilities and impel him, 
who is not already qualified with such useful knowledge, 110 
to become familiar as soon as possible with the various 
methods of demonstrating the tubercle bacillus. 
The outlay of less than fifty dollars will fully equip 
him with the necessary microscopic outfit, and the 
devotion of a few hours’ time under the direction of a 
competent instructor will do the rest. 
The medical student who does not possess sufficient 
microscopic technique to demonstrate the tubercle 
bacillus ought not to be commissioned, either by a 
medical college or the State, to practise medicine. No 
part of his preparation for accurate diagnosis is of so 
much importance as the ability to detect, by means of 
the microscope, tuberculosis of the lungs in its incipi- 
ency. It were far better he should, when engaged in 
practice, be without horse and carriage, or the display 
of surgical instruments and attractive office furniture— 
particularly the Gynecological Chair, representing 
“The Great Medical Error of the Day”—than with- 
out a good microscope and the ability to use it success- 
fully. 
The immense waste of health and life, and treasure 
also, from lack of such knowledge and technique on the 
part of physicians generally is incalculable ; for the 
statement cannot be gainsaid that tubercular consump- 
tion, if discovered and properly treated in its incipi- 
ency, is a manageable disease in numberless instances. 
I make this statement with as much confidence as I have 
in any truth in medicine. I could recount scores of 
cases, if it were necessary, in which the early recogni- 
MEDICAL MICROSCOPY. BACTERIOLOGY. 
111 
tion of the disease, by the discovery of the bacillus 
tuberculosis in the sputum, was the saving of the patient; 
for my experience has never failed me in a single in- 
stance that the presence of this bacillus in the sputum is 
the proof of the existence of either pulmonary or laryn- 
geal tuberculosis. On the other hand, the absence of the 
microorganism in the sputum is the full warrant—no 
matter how well pronounced the resemblance of symp- 
toms to favor the diagnosis of tuberculosis—that the 
case is not one of tubercular consumption. 
If all cases called “consumption” could be sub- 
jected to the microscopic test—to prove the presence or 
absence of the Bacillus tuberculosis in the sputum—the 
ghouls in human shape who now fatten from the sale 
of vile nostrums which they offer as cures for con- 
sumption, would find fewer victims to rob and feed 
upon by false promises and mirabile dictu newspaper 
stories. 
The Bacillus tuberculosis is a fine rod with clearly 
rounded ends, varying in length from 2-8 ji, but hav- 
ing an average nearly equal to the diameter of a human 
blood-corpuscle ; either straight, curved, or crooked at 
all angles, but mostly of fiddle-bow curve; lying singly 
and in pairs, sometimes overlapping each other, or 
lying in bundles or heaps, occupying both epithelioid 
and giant-cells, and strictly parasitic in its character. In 
giant-cells, the position of the bacilli is quite character- 
istic ; and by their presence a series of most interesting 
changes take place in the center of the cell, finally end- 112 
MEDICAL MICROSCOPY. 
ing in the massing of necrotic or dead cells which can- 
not be stained differentially. 
The bacilli are destitute of motion,—probably multiply 
by spores, though this mode is not necessary for perpetu- 
ation of the species, —aerobic, very selective of culture 
media, and restricted in development to narrow ranges 
of temperature with 370 C., or blood-heat, for the 
standard of most thrifty growth. In sputum espe- 
Fig. 18. 
Bacillus Tuberculosis in Sputum. 
dally, because of the thick albuminous character of the 
material, the bacterium is capable of living in a dried 
condition for several months. This fact is terribly 
suggestive of the danger of infection, particularly in 
towns and cities, from inhaling particles of dried 
sputum swept by the winds through the streets. Very 
many times, on seeing a suspected sputum on the pave- 
ment, have I picked up such a specimen for micro- BACTERIOLOGY. 
113 
scopic examination and found it loaded with tubercle 
bacilli.* 
When one’s “standard vitality” is lost from the 
effects of cold or other cause, producing a favorable 
culture-surface in the bronchial tubes, the inhalation 
and lodgment of the bacillus is not only possible, but, 
no doubt, in that way the disease is frequently con- 
tracted, as well as by breathing the air and dust of a 
hospital ward for tuberculous subjects, or the confined 
atmosphere of a private room occupied by such a 
patient. 
I think my observation fully warrants the statement 
that the sputum becomes more and more loaded with 
tubercle bacilli as the disease advances to a fatal termi- 
* There can no longer be any doubt concerning the unity of 
scrofula, lupus, and tuberculosis, or that they are due to one and 
the same microorganism. In 1887 I had the opportunity of clearly 
demonstrating not only the presence of giant-cells in lupus tissue, 
but also swarms of tubercle bacilli. (The Med. News, Phila., 
1887.) 
In searching for diseases kindred to tubercle, chronic glanders, 
syphilis, leprosy, and antinomycosis cannot escape particular atten- 
tion. They are all of bacillary origin, and produce tissue-growths 
that are strikingly similar in histo-pathologic characters. The 
closest resemblances are to be found between tubercle, syphilis, 
and leprosy. 
Indeed, so closely does the Bacillus leprce resemble the Bacillus 
tuberculosis that the most authoritative bacteriologists have been 
unable to mark much difference between them. Baumgarten has 
asserted that, both in their morphology and staining receptivity, they 
cannot be distinguished. 114 
MEDICAL MICROSCOPY. 
nation, and that their number diminishes with any im- 
provement of the patient’s condition. 
The infectious activity of this parasite defies all tem- 
peratures reaching near the boiling point; neither is its 
vitality in the least diminished by decomposition of 
animal tissues and secretions, nor contact with the gas- 
tric juices. 
Here, again, as with anthrax, the killing influence of 
direct sunlight on bacteria has another example. Koch 
found that the tubercle bacillus is killed on exposure 
varying from some minutes to several hours to direct 
sunlight, while diffuse daylight required from five to 
seven days to destroy it. 
Observations have already been made by Marshall 
Ward and others on the effect of electric arc light as a 
disinfecting agent, and with sufficient favorable result 
to encourage further experimental inquiry. 
The well ascertained thermal death-point of the 
tubercle bacillus is 70° C. for four or five minutes. 
“ The following is Kitasato’s method of preparing 
pure cultures from sputum : The patient is requested 
to evacuate his morning sputum into a sterilized double 
capsule. A flake is isolated with sterilized instruments, 
and carefully washed in at least ten watch-glasses full of 
distilled water, one after the other, to remove the 
bacteria taken up in passing through the cavity of 
the mouth. The flake is now transferred to glycerin- 
agar or serum. After it has been kept for some four- 
teen days in the incubator the first colonies form, ap- 
pearing as circular, pure, white, transparent specks, BACTERIOLOGY. 
115 
which project above the surface of the medium.” 
{Schenk.') 
Another and perhaps more simple method (Pastor’s) 
is as follows :— 
“ A patient is chosen whose sputum is very rich in 
bacilli and shows comparatively little contamination 
with other microorganisms, and he is made to rinse out 
his mouth and pharyngeal cavity repeatedly with dis- 
tilled water, and then to expectorate into a sterilized 
test-glass. The sputum, or more properly the liquid 
contents of the pulmonary cavities, is shaken up with 
sterilized water and filtered through fine gauze to re- 
move the coarser particles. A few drops of the filtrate 
are mixed with melted nutrient gelatin in such a man- 
ner as not to render it very turbid, and the mixture is 
poured out on plates which are left under bell-glasses at 
room temperature. In from three to four days the 
various colonies of bacteria contaminating the sputum 
form.” {Ibid.) 
As a matter of common observation, and an im- 
portant point of difference between a tubercular sputum 
and one which is not tubercular, is that the former when 
kept at room temperature soon becomes liquid and 
putrid, of greenish color, and exhales a most offensive 
odor, while the latter—no matter from what other form 
of chronic pulmonary disease—soon thickens by drying, 
becomes pasty, and is not offensive to the smell. 
Tubercle bacilli cannot be seen in an unstained spu- 
tum ; hence the necessity of certain staining methods. 
For the demonstration of the tubercle bacillus, either 116 
MEDICAL MICROSCOPY. 
in cover-glass or tissue preparation, I have for the last 
several years employed, to the exclusion of all other 
formula with which I am acquainted, a compound stain 
constructed on the basis of the Gibbes’ Double Stain 
(No. 16). With this “ Carbolized Double Stain”—as 
I have been in the habit of calling it,—I have been able 
to get better results, a sharper differential field, and more 
durable picture, than from the use of any other formula. 
The following is the method of employment:— 
A cover-glass having been prepared in the usual 
manner with the sputum—thinly-coated, dried, and 
several times whipped through the flame of a spirit 
lamp,—a small quantity of the stain is filtered into a 
test-tube, which should then be moved back and forth 
through the lamp flame until the fluid begins to steam. 
The cover-glass is then placed on a piece of blotting 
paper and the hot stain poured upon the coated or 
sputum surface,—just enough of the stain, drop by drop, 
to flood the cover almost to running over. It is now 
covered with a bell-glass and thus stained for five or ten 
minutes. When ready for the next step, it is picked up 
from the blotting paper with the forceps, washed, and 
sufficiently decolorized in alcohol. It is then taken 
between the thumb and forefinger of the left hand and 
wiped on the under surface, and otherwise handled as 
directed on a preceding page. 
If it be a section-mount, the slide containing the ad- 
herent section—duly prepared as described on a preced- 
ing page—is placed on a blotting paper and the same 
hot stain poured upon the section, in sufficient quantity BACTERIOLOGY. 
117 
to cover it completely. The slide is then covered with 
a bell-glass, and so kept for fifteen or twenty minutes, 
after which the right-hand end is seized with the forceps, 
the slide tipped to drain off the surplus staining fluid, 
and then washed and decolorized in alcohol,—from one 
beaker or bottle into another, until the alcohol flows off 
the slide without color. 
It is then quickly wiped with a linen cloth, free from 
lint, after which—holding the slide by the upper left 
hand corner, between the thumb and forefinger of the 
left hand—the section is flooded with absolute alcohol, 
which is poured off and on several times. After hav- 
ing been thus dehydrated, the section is next flooded 
with xylol—again and again, until it is well cleared up, 
—then the slide is wiped all around the section, a drop 
of balsam and xylol applied, and the cover-glass laid on. 
If benzol is used instead of xylol as a clearing agent, 
the stained bacilli soon fade. Another important point 
worth remembering in connection with benzol is that 
it cannot be used successfully as a clearing agent in a 
humid atmosphere. At such times, when poured upon 
the section, it produces a milky appearance. When- 
ever that occurs, xylol should be used in place of ben- 
zol. 
Croupous Pneumonia.—Two very different micro- 
organisms, with some features in common, have been 
found in the lungs and sputum of patients suffering 
from this disease,—the Micrococcus pasteuri, discovered 
by Sternberg in 1880, and the Pneumobacillus of Fried- 
laender, both varieties, presumably, being of a specific 118 
MEDICAL MICROSCOPY. 
character. The relative frequency, however, of the de- 
tection of the two forms in a given number of cases 
differs widely. Weichselbaume found the “ Diplo- 
coccus ” in ninety-one cases out of a hundred, while, on 
the other hand, in one hundred cases the pneumobacillus 
was only present nine times. 
Fig. 19. 
Recent Croupous Pneumonia, a. Alveolar Septa with Injected 
Capillary Vessels, b. The Exudation. 1-300. 
Though never presenting as distinctly rod-shaped as 
Friedlaender’s bacterium, Sternberg’s Micrococcus pas- 
teuri is sufficiently well marked to distinguish it from 
all other microorganisms. Both of them develop a BACTERIOLOGY. 
119 
capsule in the animal body and never outside of it, and 
may be artificially cultivated, but the culture-medium 
must have at least a faint alkaline reaction, as an acid 
renders growth impossible. 
Both varieties are alike immotile, semi-anaerobic, 
and may be found in the mouth and nasal passages of 
healthy persons, also in the lungs and expectorated 
matter of patients suffering from other pulmonary dis- 
Fig. 20. 
Pneumonococci. (Jaksch.) 
eases. The thermal death-point for this microorganism 
is 56° C. 
The strongest differential point is, probably, that 
while Sternberg’s Micrococcus pasteuri may be beau- 
tifully stained and demonstrated by Gram’s method, 
the pneumobacillus of Friedlaender is decolorized and 
undemonstrable. Sections showing fibrin-choked al- 
veoli may be prepared either with Weigert’s Fibrin 120 
MEDICAL MICROSCOPY. 
Stain, or the author’s Safranine method. (See Nos. 
23-24.) 
The general knowledge on the subject concerning the 
causal relation of these microorganisms to croupous 
pneumonia accepts the Micrococcus pasteuri as being 
the true factor of the disease. 
Diphtheria.—The microorganism of diphtheria is 
now well known as the Klebs-Loffler bacillus. The 
rods are about the same length as the tubercle bacillus, 
either straight or curved, crooked or club-shaped, and 
at least twice as thick as the tubercle bacillus. Some of 
the rodlets appear much longer than others, but the 
Fig. 21. 
Bacilli of Diphtheria. X 1000. (Gould.) 
long ones are found on careful inspection to be made up 
of single pieces linked so closely together by their ends 
as to give the appearance of being a continuous body. 
They are motionless and have no spores. The false 
membrane is a network of fibrin enclosing in the meshes 
epithelial cells, blood and pus-corpuscles, and various 
microorganisms, besides the Klebs-Loffler bacillus which 
is found in all typical cases of the disease and retains 
its virulence, even when completely dried, for several 
months. The older and thicker parts of the false 
membrane are the sites of the most luxuriant growth 
of the bacillus, but it may be grown outside the animal BACTERIOLOGY. 
121 
body, on gelatin and in blood serum. It is semi-anae- 
robic and thrives best at a temperature between 20° 
to 420 C. 
The method of staining may be made a means of 
differential diagnosis, Loffler’s alkaline methyl-blue giv- 
ing good demonstration with deeply colored ends of the 
bacillus, while Gram’s method yields a negative result. 
Dr. Sims Woodhead’s method of staining the micro- 
organism is as follows: It is only necessary to remove 
a small fragment of the false membrane by means of a 
bit of absorbent cotton and carried with the forceps to 
a cover-glass, on which it is thinly spread out, then 
dried and heated over the flame of a spirit lamp in 
the same way as a sputum preparation, after which a 
drop of Loffler’s solution is placed on the slide and the 
cover-glass laid on it. Superfluous fluid should be re- 
moved with a piece of blotting-paper and the mount 
examined at once. The organisms occur in small 
groups, and, if present, may be easily discovered. 
Scarlet Fever.—The bacteriologic study of scar- 
let fever has not yet resulted in the discovery of the 
specific germ to which the disease is due. Various 
investigators have discovered cocci in typical scarla- 
tina, but the organisms in no way could be distin- 
guished from the Streptococcus pyogenes derived from 
pus. Very recently Dr. Klein succeeded in separating 
a microorganism from the blood which he regards as 
the true virus or contagium of the disease, notwith- 
standing its close resemblance to the Streptococcus 
pyogenes, but the experiments upon which he based his 122 
MEDICAL MICROSCOPY. 
claim have not been accepted without serious question. 
Neither has the very able experimental work of Jamie- 
son and Edington, reported in the British Medical 
Journal, 1887, fulfilled the hope it so brilliantly 
encouraged. So, at present, it may be here stated that 
up to this time no microorganism has yet been dis- 
covered and isolated which can confidently be accepted 
as having causal relation to scarlet fever. 
Fig. 22. 
Gonococci contained in Pus-cells. (From Friedlaender.) 
Gonorrhea.—The doctrine of the causal relation 
of the microorganism called Go?iococcus—first dis- 
covered in 1879 by Neisser, of Breslau—to gonorrhea 
is now generally accepted. The very infectious nature 
of gonorrheal pus distinguishes it from all other pus- 
products. 
The cocci, about 0.83 ;j. in diameter, are arranged 
singly, in pairs, and in groups, often closely set BACTERIOLOGY. 
123 
against each other in the interior of the pus-cells sur- 
rounding the nuclei, and can be easily identified. 
Cultivation experiments have been uniformly unsuc- 
cessful with all media except human blood-serum, and 
even in this the growth is very slow, the most favorable 
range being between 330 to 370 C. The death-point 
is 6o° C. 
The cocci increase by fission at right angles, aerobic, 
and are easily stained with the “ Carbolized Double 
Stain.” They do not respond to Gram’s method. 
Cover-glass preparations should be handled in the 
same manner precisely as directed for tuberculous 
sputum mounts. 
Syphilis.—Lustgarten found small comma-bacilli, 
curved and somewhat resembling the tubercle bacillus, 
in the discharge of the primary lesion of syphilis, also 
in the gummata of the tertiary form of the disease, but 
the microorganism—the alleged Bacillus of syphilis— 
has not yet been successfully cultivated outside the 
animal body. Indeed, its pathogenic character has 
not been accepted without question as having true 
causal relation to syphilis. 
While the presence of the Lustgarten microorganism 
affords a valuable indication of the disease, extreme 
caution should be exercised to prevent confounding it 
with the Smegma bacillus found in normal smegma prce- 
putiale and vulvare, which it so much resembles, both 
in appearance and staining reaction. 
Kamen has reported the case of a child in whose 
sputum he found the Lustgarten bacillus. 124 
MEDICAL MICROSCOPY. 
The two forms of bacilli may be easily differentiated 
by the alcohol test. After staining, the syphilis bacillus 
parts with the stain very slowly in alcohol : the smegma 
bacillus soon becomes paled and a blank in alcohol. 
The syphilis bacillus, so-called, may be stained with 
a solution of gentian violet in five per cent, carbolized 
water, used hot for both cover-glass and sections : then, 
after staining for a couple of hours, to go into absolute 
alcohol; then into one and a half per cent, solution of 
permanganate of potassium, which deprives the tissues of 
their color while the bacilli retain their violet tint. To 
Fig. 23. 
Bacilli of Syphilis. X 1000. (Gould.) 
remove the precipitate of manganese which forms upon 
the specimens, they are immersed in an aqueous solution 
of sulphurous acid. This is “ Ehrlich’s Solution ” with 
carbolized water instead of anilin water. 
A far more simple and equally effective method may 
be accomplished with Lofifler’s methyl-blue poured on 
hot, decolorized with water, and then double stained 
with a watery solution of safranin. 
Glanders.—The frequent occurrence of glanders in 
the human body is sufficient excuse for the mention of 
the disease in these pages. The specific microorganism 
is a slender bacillus, with rounded ends. Baumgarten BACTERIOLOGY. 
125 
and Rosenthal claim that it is spore-producing, as shown 
by means of double staining ; but it requires further 
observation to settle the question. It is from 2 to 5 [j. 
long and 0.5 to broad, or about the size of the 
tubercle bacillus, but a little thicker. It was first dis- 
covered by Loffler and Schiitz, and may be found in the 
nodules and ulcers which form in the nasal passages and 
neighboring lymphatic glands, also, sometimes, in the 
blood and internal organs. 
Fig. 24. 
Bacillus of Glanders. ( Jaksch.) 
In a dried state it may retain its infective power for 
several months. It grows in various media,—agar-agar, 
glycerin-agar, and on potatoes, at a temperature between 
30° and 40° C. It does not thrive well on gelatin, be- 
cause its development is suspended at a temperature 
above 420 C. It is very motile. Its thermal death- 
point is 550 C. 
Cover-glass preparations may be made of the pus-dis- 
charge in the usual way, and stained either with “ Car- 126 
bolized Double Stain ” or Loffler’s Solution. Sections 
should be dealt with as directed for mounting enteric or 
typhoid fever tissues,—stained in the same way, and 
dehydrated with benzol instead of alcohol. 
Leprosy.—The microorganism of leprosy is a slen- 
der rod with somewhat pointed ends, and probably 
not quite as long as the tubercle bacillus, its length 
being, upon an average, about two-thirds the diameter 
of a human blood corpuscle, or 4-6 ;j. long, and less than 
1 n wide. It contains two or three spores, and, in tissue, 
presents a beaded appearance. The bacilli are immo- 
MEDICAL MICROSCOPY. 
Fig. 25 
Bacillus lepra. X 1200. (Gould.) 
Bacillus of Leprosy. X 800 Diam. 
(Coplin.) 
tile, and principally distinguished from tubercle bacilli 
by staining readily in aqueous solutions of the anilin 
dyes. Beautiful preparations may be made with the 
carbolized double stain (No. 16). 
The specific bacillus is found in all leprous processes 
and products, in the cutaneous connective tissue, in the 
spleen, kidneys, liver, testicles, and lymphatic glands, 
but not in the blood. Cultivation-experiments have 
not been generally successful. It thrives best at a tem- 
perature from 370 to 38° C., and is very sensitive to 
some of the basic anilin colors. BACTERIOLOGY. 
127 
Tetanus.—The importance of this disease of wound- 
infection, with a clinical history distinguished from all 
other affections, makes necessary its brief description. 
The specific microorganism is a very slender rod, of 
a peculiar “ pin-shape,” having a spore at one end 
which gives the body a drumstick appearance. It was 
discovered by Nicolaier in pus from wounds of persons 
who had died of the disease, and in garden mold. 
It has also been found in old, crumbling walls, putre- 
scent fluids, and other decaying substances, including 
Fig. 26. 
Bacillus tetani. X 3oo Diam. (Coplin.) 
manure heaps. The discovery of the last-named source 
no doubt gave origin to the opinion of some French in- 
vestigators that the disease develops in persons who fre- 
quent stables or are in contact with horses. 
Many authorities believe that tetanus is an infectious 
disease and that the pin-shaped bacillus is the factor of 
its communicability. Brieger and Nissen have experi- 
mentally demonstrated the presence of several ptomains 
in the blood, with which they have induced tetanic 
symptoms and poisoning in various animals. To de- 128 
MEDICAL MICROSCOPY. 
stroy the spores requires exposure for five minutes, at 
least, to the action of steam at ioo° C. It is slightly 
motile, and can withstand a temperature of 8o° C. 
without losing its pathogenic power ; grows in various 
culture-media and liquefies gelatin; rigidly anaerobic 
and thrives best at a temperature from 36° to 38° C. 
It colors equally well with several of the anilin dyes, 
and may be demonstrated by Gram’s method. Here, 
also, the “ Carbolized Double Stain ” maybe employed 
with very satisfactory effect (No. 16). 
Influenza.—The alleged discovery of the bacillus 
of influenza by Pfeiffer and confirmed by Kitasato, 
Fig. 27. 
Bacilli of Influenza. X 1000. (Gould.) 
Canon, and Babes, is probably the very latest addition 
to the list of pathogenic bacteria cultivated outside the 
animal body. The disease is of such universal impor- 
tance—having invaded whole continents within the last 
several years, and is now, at the beginning of the year 
1894, wide-spread in its visitation in the United States 
and over Europe—that we may confidently expect very 
soon more exact knowledge on the subject of the bac- 
teriology of the disease. 
The influenza bacillus is the smallest of the rod bac- 
teria, very slender, and not thicker than the bacillus ot BACTERIOLOGY. 
129 
mouse-septicemia. Kitasato succeeded in cultivating it 
to the fifth generation in glycerin-agar. Canon found 
the same bacterium in the blood of patients suffering 
from the disease, and cultivated it from that source. 
The diminutive rods occupy for the most part, the white 
corpuscles, and may be stained with Chenzynski’s solu- 
tion of methyl-blue and eosin (No. 5). By this pro- 
cess the red blood-corpuscles are stained red, the white 
corpuscles and bacilli blue. 
Anthrax.—Although first thoroughly studied by 
Fig. 28. 
Fig. 29. 
Bacillus anthracis. X 800 Diam. 
{Copi in.) 
Bacillus of Symptomatic Anthrax, Flagel- 
late Form. X 1000. {Gould.) 
Koch, the Bacillus anthracis was seen by Pollender in 
the blood of animals suffering from the disease as early 
as 1849. It ts found in the blood and tissues of animals 
dying or dead of the disease. Pasteur made the dis- 
covery that it is spread by earth-worms ; but the state- 
ment has not been confirmed by other observers. The 
rods are arranged in chains of varying length, from 
3 to 20 /i long, and x.o to 1.25 /j. broad, and remark- 
able for their evenness of outline and squarely cut 
ends. 
They are immotile, aerobic, and may be grown in 130 
MEDICAL MICROSCOPY. 
gelatin (which it liquefies), agar-agar, on potatoes, and 
blood serum. The multiplication is by egg-shaped 
spores, springing from the long axes of the maternal 
cells, the thermal death-point of which is ioo° C. 
The Bacillus anthracis meets every experimental re- 
quirement to prove that it is the cause of the disease, 
and is one of the best examples of a disease due to 
microorganisms. 
For the demonstration of the bacilli in sections, the 
“ carbolized double stain ” maybe used successfully. 
Very beautiful preparations also may be made according 
to Gram’s method. 
Amoeba coli—Amoeba dysenteries.—This protozoon 
Fig. 30, 
Amoeba coli. 
has been invested with so much interest since the studies 
of Councilman and Lafleur, Osier and Dock, that a 
brief description of its character should not be passed. 
Losch was the first to connect this protozoon with 
dysentery in a causal relation, and his observations 
have been fully confirmed by the distinguished American 
authorities just named. It may be described as follows : 
Cellular body of circular form having a diameter of 20 BACTERIOLOGY. 
131 
to 35 [i contractile, consisting of a hyaline and coarsely 
granular protoplasm, with a round nucleus and hyaline 
vesicle, but no cell wall. 
According to the observations of Councilman, “ the 
amoeba when at rest are round or slightly oblong bodies, 
consisting of an outer pale, homogeneous substance en- 
closing a somewhat greenish, highly refractive mass, 
which contains vacuoles of various sizes and a nucleus. 
Movement is their distinctive feature, however, and con- 
sists, first, of a progressive motion, and, secondly, of a 
protrusion and withdrawal of pseudopodia, both of 
which vary in activity. The pseudopodia are formed 
from the outer homogeneous part, which may, how- 
ever, be otherwise invisible both in the resting and 
moving state. The amoeba often contain foreign bodies, 
such as red corpuscles, pus cells, blood pigment, micro- 
cocci, bacilli, and their spores. 
“ Entering probably with the food, they pass on 
until the large intestine is reached, where the alkalinity 
necessary to their growth is obtained. Here they pene- 
trate and undermine the mucous membrane, producing 
their effect by liquefying the tissues, and thus causing 
ulceration and necrosis. In the mucous membrane 
they are found chiefly in the submucosa, in the lymph- 
spaces and blood-vessels, and in the gelatinous con- 
tents of the ulcers. They may penetrate to the liver 
either by the portal vessels or through the peritoneum— 
sometimes causing peritonitis,—and set up abscesses 
which in the former case may be multiple, in the latter 
lie close to the surface of the right lobe, the commonest 132 
MEDICAL MICROSCOPY. 
position.” According to Councilman and Lafleur, the 
liver shows no inflammatory reaction, and the abscess- 
cavity is filled, not with pus, but with debris of liver 
tissue, or sometimes a necrosed mass ; the contents may, 
however, be old pus, the suppuration being due to the 
action of microorganisms conveyed by the amoeba (Kar- 
tulis). The liver abscess may extend directly so as to in- 
volve the lung, or the amoeba may traverse the diaphragm 
and set up an abscess by liquefying the tissue; but here 
the cavity is surrounded by an area of interstitial in- 
flammation. That the microorganisms do not travel 
by the lymphatics is shown by their absence from the 
mesenteric glands, and they have no special preference 
for the lymphoid follicles. 
“ Ante-mortem they may be found in the stools, par- 
ticularly the gelatinous particles, in the pus from liver 
abscesses, and in the sputum in cases of abscess of the 
lungs, and may be examined in the fresh state, best on 
the warm stage or in cover-glass preparations, or sections 
may be cut from portions of feces hardened and embed- 
ded in celloidin. Councilman and Lafleur obtained the 
best results with sections of tissue hardened in alcohol and 
stained in Ldffler’s methyl-blue, by which method the 
amoeba are colored dark-blue, but unevenly. The 
nuclei are best brought out by hardening in Flemming’s 
solution. 
“ Kartulis succeeded in obtaining pure cultures by 
inoculating alkaline infusion of straw with some of the 
contents of a liver abscess in which no bacteria were 
present.” (Dawson.) BACTERIOLOGY. 
133 
Plasmodium malariae (.Polymitus malaria, Dani- 
lewsky). The first successful step toward the present 
exact knowledge of malarial intoxication was made by 
Laveran in 1880, whose name is indissolubly connected 
with the affection. From that well-established starting 
point, Marchiafava and Celli began their brilliant in- 
vestigations which resulted in the discovery of the par- 
ticular parasite, or hematozoon, which is now accepted 
with remarkable unanimity of medical opinion as the 
determining cause of the disease, and generally believed 
to stand in close relation to soil. 
In this country the careful studies of Councilman, 
Abbot, Sternberg and Osier, have abundantly con- 
firmed the descriptions furnished by Laveran, Dani- 
lewsky, Marchiafava, Celli, Golgi, Guarnieri, Crook- 
shank, and other distinguished foreign observers, and 
given the subject a practical turn of the very highest 
clinical importance from the stand-point of differential 
diagnosis. 
As indicated by Laveran, there are several develop- 
mental or evolutionary processes which successively take 
place in the life-history of the micro-parasite, giving a 
variety of body or shape corresponding with the well- 
recognized three stages of the disease. The different 
phases of the hematozoon have been grouped as intra- 
corpuscular and or those within the 
red blood-corpuscle and those free in the blood serum. 
The intra-corpuscular bodies are of three forms 
or kinds: — 
“ First, structureless protoplasmic bodies, much 134 
MEDICAL MICROSCOPY. 
smaller than, and within or attached to, the red blood- 
corpuscles (Fig. 31). These rapidly change their shape, . 
exhibiting ameboid movement,” and were named by 
Marchiafava and Celli Plasmodium malaria. 
“ Second, minute masses of finely granular or of hya- 
line protoplasm enclosing granules of pigment (Fig. 
31). These forms are sometimes present in large num- 
bers, and at other times can be found only with difficulty. 
Fig. 31. 
Non.pigmented Amoeboid Forms. 
Pigmented Amoeboid Forms. 
They are more or less spherical, but exhibit ameboid 
movement, and rapidly change their form. The pig- 
ment granules are also in active movement. There may 
be one or more of these ameboid bodies to a blood- 
corpuscle, and they vary in size ; one may occupy the 
whole of the corpuscle. In cases of pernicious malaria 
similar bodies may be seen in tissue sections, in the cor- 
puscles filling the capillaries. 
“ Third, forms which appear like isolated grains, and larger homogeneous bodies surrounded by clear spaces 
which change in outline. 
“ Extra-corpuscular Bodies.—These are the most 
striking and perhaps the most interesting forms. 
First, the semi-lunar bodies of Laveran. These are 
crescent-shaped bodies, sometimes pointed at the 
extremities, but more usually rounded off (Fig. 32). 
They are not always curved ; some, indeed, are almost 
spherical, and others sausage-shaped. They are 
motionless. In many specimens a delicate line is 
visible on the concave side of the crescent, connecting 
BACTERIOLOGY. 
135 
Fig. 32. 
Plasmodium malariae (Extra-corpuscular Bodies). 
the extremities. On careful examination this is found to 
be the edge of a very delicate membrane. The body 
is composed of homogeneous protoplasm. Centrally 
placed is a collection of pigment granules, which on 
careful examination can be distinctly seen to be in move- 
ment. The semi-lunar bodies vary in number in different 
cases. Sometimes several can be seen in the field at the 
same time, and in other cases they are only observed 
after a long and patient search. They are, as a rule, 
free in the serum ; but they have also been seen within 
the red blood corpuscles. 136 
MEDICAL MICROSCOPY. 
“ Second, finely granular masses of protoplasm, which 
arise, according to Golgi, from the intra-corpuscular 
pigmented bodies. The pigment is collected in a rosette, 
and the protoplasm by segmentation gives rise to a 
number of small spherical forms, which are ultimately 
set free (Fig. 33). Golgi believes that these changes 
Fig. 33. 
Rosette Forms with Segmentation. 
Fig. 34. 
Flagellated Forms, i. A flagellated spherule; (a) the same in the interior 
of a phagocyte; (£) free motile filaments. 
occur in definite relation to the development of the 
paroxysm. 
“ Third, spherical, pear-shaped, or ovoid bodies, rather 
smaller than the red blood corpuscles, and provided with 
one or more actively motile flagella (Fig. 34). These BACTERIOLOGY. 
137 
flagella are long, lash-like filaments, which by their 
activity set the neighboring blood corpuscles in motion. 
Free filaments in active movement have also been 
observed. 
Fourth, small spherical pigmented bodies about one 
quarter the size of a red blood corpuscle, which exhibit 
ameboid movement.” (Crookshank.) 
The process of segmentation and multiplication, ac- 
cording to Golgi, covers a period from two to three 
days. At first the new plasmodia adhere to the edge of 
the blood-corpuscle, next become free, and subsequently 
invade other corpuscles. 
And thus it is seen that by a proper examination of 
the blood it is possible to distinguish malaria with abso- 
lute certainty from all other affections which closely 
resemble it, or cases of obscure sepsis, endocarditis, and 
tuberculosis. 
Artificial cultivation of the malarial parasite has not 
been successful, neither has its pathogenic character 
been established, notwithstanding the assertion made 
by Marchiafava and Celli that inoculation of a healthy 
person with blood containing the parasite will produce 
a paroxysm with development of the specific hemato- 
zoon. 
Staining.—Various methods may be employed for 
the demonstration of the malarial parasite. To make a 
permanent preparation, a very thin layer of blood should 
be dried on either the slide or cover-glass in the usual 
way, and stained with Plehn’s solution (No. 6). By 
this method the red corpuscles appear a light red, leu- 138 
MEDICAL MICROSCOPY. 
kocytes light blue, and their nuclei a deep blue, the 
eosinophil granules of the leukocytes a deep red, while 
the parasites are stained blue. 
Malachowski’s method is very easy to follow. He 
lays the cover-glass preparation in alcohol and stains it 
in a mixture of equal parts of a one per cent, solution of 
eosin and dilute aqueous solution of borax and methyl- 
blue (No. 31), by which method the red corpuscles 
show yellowish-red, the nuclei of the leukocytes violet, 
and the plasmodium blue. 
Fig. 35. 
Spirillum of Relapsing Fever. X 800 Diam. (Coplin ) 
For immediate diagnosis the method recommended 
by Jaksch is of great value. Methylene-blue is dis- 
solved in normal (0.6 per cent.) salt solution until the 
fluid is deeply colored, which is then filtered, sterilized, 
and sealed in small quantities in thoroughly sterilized 
test-tubes. 
The point of the finger is carefully cleansed, a drop 
of the staining fluid applied, and through this drop the 
finger is pricked with a needle. The flowing blood 
thus mixed with the stain is spread as thinly and quickly 
as possible upon either the slide or the covering-glass, BACTERIOLOGY. 
139 
and examined with a 1-12 oil-immersion lens and Abbe 
condenser. Evaporation may be prevented by sealing 
the edge of the cover-glass with paraffin. 
Actinomycosis.—The wide distribution of this 
disease of cattle, principally, and the increasing num- 
ber of cases of the same affection in human beings 
from accidental inoculation with the specific fungus in 
the pus-product developed in a lower animal, has made 
the affection a matter of so much public concern that 
the general knowledge on the subject of the exact 
Fig. 36. 
Actinomyces (Ray Fungus). X 800 Diam. (Coplin.) 
nature of the disease has been greatly improved within 
the last ten or twelve years; so that now no longer any 
doubt exists concerning its communicable character 
from animal to animal and from animal to man. 
The fungus character of the disease was discovered as 
early as 1845 by Langenbeck, but it was not until 1877 
that the specific parasite—Actinomyces bovis—was dis- 
covered and isolated by Bollinger. 
Following Bollinger’s discovery of the parasitic nature 140 
MEDICAL MICROSCOPY. 
of the disease as it occurs in cattle and hogs, Ponfick 
and Israel furnished the proof of its easy communica- 
bility to man, and since the publication of their careful 
investigations many such examples of transmission from 
cattle to man have been reported. 
The disease has long been familiarly known to stock- 
growers as “big-head,” “big-jaw,” or “ lumpy-jaw,” 
and, in surgical parlance, medullary sarcoma, osteo-sar- 
coma, etc. The presence of the seed-like granules in 
bovine pus is sufficient to make sure the diagnosis, even 
without the aid of the microscope. 
That the disease “ lumpy-jaw,” or actinomycosis, in 
cattle as it is seen in this country, and fully described 
by Dr. D. A. Salmon, Chief of the Bureau of Animal 
Industry (see eighth and ninth annual reports, 1893), is 
the same affection described by Bollinger, has been 
clearly shown by the distinguished studies of Dr. W. T. 
Belfield, of Chicago, to whom belongs the credit of 
showing the unity, beyond all question. 
In cattle, the lower jaw is the usual site of the dis- 
ease, and the infectious fungus is generally believed to 
effect entrance by the mouth and in some manner find 
lodgment either in a carious tooth or on an abraded 
gum. But no matter what the mode of introduction of 
the pathogenic microorganism, a focus of inflammation 
is soon established and a nodular growth, composed 
chiefly of round cells, granulation tissue, and fibrous 
bands, follows in more or less active order. These 
infective nodules are of various sizes according to the 
age or luxuriance of growth, but soon break down in BACTERIOLOGY. 
141 
suppuration and result in pus-cavities. Between these 
centers of suppuration, fibrous tissue of varying degrees 
in density constitutes the structure of variously sized 
tumors which soon form and give the characteristic 
appearance. These masses may reach the size of the 
fist, or larger, in the jaw of a bullock ; in the human 
subject the soft parts of the neck, the mediastinal tis- 
sue, and the lungs are the parts most frequently in- 
volved. In some cases the histologic structure of a 
bovine specimen may so closely resemble tuberculous 
tissue that but for the central presence of the fungoid 
masses the differential diagnosis would be most difficult, 
if not impossible. This is very true, as shown in a 
specimen nodule which I have in my possession, sent 
from the Michigan Agricultural Experimental Station, 
Lansing, by Professor Grange. 
The quality of the pus and discovery in it of the seed- 
like sulphur-colored particles—constituting groups of 
Actinomyces, or the club-shaped degeneration forms of 
the parasite—will clear up the diagnosis. 
According to Israel and Wolff, the disease is classed 
with the polymorphic fission-fungi. 
Artificial cultures on glycerin-agar and potato have 
been successful in the hands of various observers. 
Whatsoever the medium employed—glycerin-agar, 
potato, meat-broth, gelatin, eggs, etc.,—masses of 
closely packed yellowish-colored granules varying in 
size, but not larger than a hempseed, are developed in 
a short time. 
Cover-glass preparations of pus may be made in the 142 
MEDICAL MICROSCOPY. 
usual manner and variously stained. Sections may be 
successfully stained by several methods—Gram’s, 
Weigert’s, Plaut’s, or Ehrlich’s. A method not hereto- 
fore published is as follows :— 
Place the slide, with section fixed, in “ Carbolized 
Hematoxylin Comp.” (No. 22) for ten minutes; then 
wash in acidulated water for two or three minutes ; then 
in plain water ; then in alcohol for five or six minutes ; 
then cover with “ Carbolized Fuchsin Comp. ” (No. 16) 
for ten minutes ; then decolorize in alcohol, clear up 
with xylol, and mount in balsam and xylol, precisely 
as directed for the demonstration of the bacillus tuber- 
culosis in tissue. 
XI. THE EXAMINATION OF 
NEOPLASMS. 
The importance of clinical microscopy finds many 
examples in the differentiation of tumors and other 
abnormal growths to determine whether the particular 
neoplasm is benign or malignant, so that the physician 
or surgeon in charge of the case may have due notice 
thereof and govern himself accordingly. 
In the practice of medicine and surgery the necessity 
of such examinations of diseased tissues comes next in 
frequency to sputum examinations, and should always, 
if possible, precede the use of the knife. Hence the 
value of the rapid method detailed on a preceding 
page of preparing a perfect and permanent mount of 
any tissue specimen for microscopic examination and THE EXAMINATION OF NEOPLASMS. 
143 
differential diagnosis, within the space of four or five 
hours from the time of its removal from the living 
body. 
A tumor in strict histo-pathologic sense represents 
a swelling caused by a new growth, and should thus be 
distinguished from a simple inflammatory swelling, the 
protrusion of an abscess, a hernia, or simple enlargement 
from hypertrophied muscular tissue. 
The classification most useful for the purposes of this 
volume is that founded upon the microscopic character 
of the neoplasm, as follows :— 
Tumors made up of normal adult human tissue, either 
entirely or in modified form, are— 
The Fibromata, 
The Lipomata, 
The Chondromata, 
The Osteomata, 
The Papillomata, 
The Adenomata, 
The Lymphomata, 
The Myomata, 
The Neuromata, 
The Angeiomata, and 
The Lymphangeiomata. 
Each of these different forms may have its subdivi- 
sions and blendings with another. 
1. The Fibromata.—These tumors may either be 
hard or soft. Hard fibrous tumors are always firm 
and encapsuled. On section, they present a glistening 
surface of grayish or pinkish color, and even to the un- 
aided eye may show the concentric arrangement of 
their component interlacing bands. They occur in 
various parts of the body, are neither painful nor tender 
on pressure—unless involving a nerve-sheath or terminal 144 
MEDICAL MICROSCOPY. 
—and in some instances it may not be easy to 
distinguish such an innocent tumor from a spindle- 
celled sarcoma. A distinctive feature of hard fibromata 
is, that though often multiple they show little or no 
tendency to return after complete removal, and but 
rarely ulcerate. They are liable to various forms of 
degeneration, the most common being partial mucoid 
softening and calcification. They are essentially 
innocent growths. 
Soft fibrous tumors are simply masses of connective 
tissue occupying submucous and subcutaneous sites, 
and, as a rule, are more or less pedunculated. The 
scalp, labia majora, scrotum, and buttocks are more fre- 
quently involved by such soft fibromata, which may 
reach great size and be multiple in character. The 
ugly growths often seen on the face and scalp, called 
wens, belong to this variety. In cases of molluscum 
fibrosum we have another example, and in this form the 
loose fibrous tissue may attain great volume, as seen in 
the immense masses that sometimes occupy the buttocks 
and other parts. These growths are often edematous 
in appearance, more or less fatty, thus resembling the 
lipomata. In this class, cheloid growths should be 
included. 
The simple polypi of the nose, ear, uterine neck, 
and meatus urinarius are variously made up of modified 
fibrous tissue, and have usually a large blood-supply, so 
that they bleed easily and sometimes profusely when 
their mucous covering is broken. In the particular 
character of the connective-tissue cells and mucous THE EXAMINATION OF NEOPLASMS. 
145 
quality of the matrix, they resemble the myxomata ; or, 
by containing in their interior the gland-structure from 
which they spring, may resemble the adenomata in 
histologic quality. 
2. The Lipomata.—These tumors are composed 
of normal adipose tissue, and are met with in all parts 
of the body in which adipose tissue is normally found ; 
they are often multiple. 
The true lipoma should not be confounded with a 
local hypertrophy of the subcutaneous fatty layer. In 
the true lipoma, the dimpling of the skin-covering when 
the tumor is moved or shaken is very characteristic. 
Such tumors are made up of variously sized fat lobules 
and involve no danger to life. Sometimes they are of 
congenital origin. In other cases, they develop as the 
result of pressure, and may become very painful. They 
vary in amount of connective tissue according to the 
size and number of lobules. Secondary changes are 
not common. In some rare cases mucoid softening 
occurs ; in others the fibrous septa may show calcareous 
degeneration. 
When removed by the knife, the operation should be 
a thorough enucleation, for otherwise recurrence is 
likely to take place. 
3. The Chondromata, or Cartilage-tumors.— 
These may be divided into those which grow in con- 
nection with bone and those which develop in the soft 
parts. When the growth is from the bone it is called 
ecchondroma ; when from a sheath of cartilage, it is an 
enchondroma. The last named variety is the simplest 146 
MEDICAL MICROSCOPY. 
form, and usually commences its growth in the fingers, 
toes, and other parts during the period of adolescence. 
The tumors, with but few exceptions, are multiple, and 
never exhibit malignancy of character. The ecchon- 
dromata belong usually to a later period of life; they 
may involve any of the bones, often attaining immense 
size, particularly in the shafts of the long bones, and 
present a high degree of malignancy. 
Cartilaginous tumors may involve the soft parts, such 
Fig. 37. 
Osteoid Chondroma. {Gould.) 
as the parotid gland, the testis, and more rarely the 
breast, the ovary, the submaxillary and lacrymal 
glands, the kidney, and even the lung; but such ex- 
amples as the last mentioned are variously mixed tumors, 
and more or less malignant according to the particular 
cell-admixture. In density, some of them are so hard 
as to be appropriately named fibro-chondromata, others 
so soft that they are denominated myxo-chondromata. 
Besides these, there are the chondro-sarcomata and 
osteo-chondromata. Softening may take place at the THE EXAMINATION OF NEOPLASMS. 
147 
center and give rise to one or more cysts; or the de- 
generation may be either that of calcification or a true 
ossification. Finally, the matrix may be hyaline or 
fibrous, with variously shaped cells. 
4. The Osteomata are tumors composed of bone, 
usually appearing in adult life, and more frequently 
involving the bones of the face than other parts. They 
have been divided to suit each phase of their growth. 
5. The Papillomata are divided as soft and moist, 
hard and dry, according to their character, which de- 
pends upon the particular structure from which they 
spring. The soft or mucous variety occurs on the lips, 
tongue, soft palate, in the larynx, about the anus, on 
the glans penis, glans clitoridis, and vulva (condylo- 
mata); at the orifice of the female urethra, in the 
uterus, vagina, bladder, and intestines. The epidermic 
papillomata are distinguished by their dense structure, 
and include warts and corns of all descriptions. 
Besides these two varieties, there is another, the 
serous and synovial growths. Many of the papillomata 
are of gonorrheal and syphilitic origin. They are made 
up of connective tissue, numerous vessels, and epithelial 
elements, exactly corresponding with the structure of 
the part from which they grow. 
6. The Adenomata of typical character are made 
up of the particular gland-structures from which they 
grow, and are non-malignant. These tumors, however, 
are always unknown quantities for the reason that in 
their growth they may suddenly depart from the benign 
type and assume malignancy. Upon this well-known 148 
MEDICAL MICROSCOPY. 
fact is based the practice of all wise surgeons of remov- 
ing such tumors, especially of the breast, with the 
knife as soon as possible after their discovery. 
Adenoma of the breast is a rounded tumor, occurring 
oftener near the axillary border than elsewhere, com- 
pletely encapsuled, and showing no tendency to recur 
after removal. On microscopic section it differs but 
little from normal mammary gland structure, and is 
frequently met with in young women, usually during 
the child-bearing period. When admixed with some 
other heteroplastic growth, the form may be an adeno- 
fibroma, adeno-myxoma, or adeno-sarcoma. The 
most frequent secondary changes are fatty degeneration 
and mucoid softening. The parts more frequently in- 
volved besides the breast are the ovary, testis, prostate, 
thyroid, parotid, glands of mucous membranes, seba- 
ceous and sweat-glands. 
7. The Lymphomata.—Tumors of this class 
most frequently occupy the cervical, submaxillary, 
axillary, and inguinal glands ; and, as a rule, are per- 
fectly innocent. In some cases they grow to enormous 
size, and thus produce great deformity. Occasionally 
they assume malignant properties, and are followed by 
secondary growths distant from the initial tumor. Mi- 
croscopically, the structure—that of an ordinary lym- 
phatic gland—differs but little from a small round-cell 
sarcoma. 
8. The Myomata.—Just as there are two kinds of 
muscle, so there are two forms of muscle-tissue tumors : 
the leiomyoma, representing the smooth or unstriped muscular fiber, and the labdo- or rhabdomyoma, re- 
presenting striped muscular fiber. The wall of the 
uterus, either on the external or internal surface, is a 
favorite site for tumors of the smooth or unstriped 
variety, which are often pedunculated. 
In some congenital tumors of the heart and kidney, 
striped muscular fiber—the labdo- or rhabdomyoma— 
has been found. 
The secondary change most likely to take place with 
THE EXAMINATION OF NEOPLASMS. 
149 
Fig. 38. 
Leiomyoma of the Uterus. (Gould.') 
the myomata is calcification. Now and then mucoid 
softening and the formation of cysts, accompanied with 
more or less hemorrhage, are met with. These tumors 
are always innocent. 
9. The Neuromata.—These painful tumors are, 
fortunately, very rare. In the majority of cases they 
cannot be easily distinguished from fibromata, because 
of their immediate contact with nerve fibers. They are 
of small size and slow growth, hard, generally multiple, 
and confined to the branches of some particular periph- 150 
MEDICAL MICROSCOPY. 
eral nerve. In the traumatic variety—the amputation- 
neuromata—the tumor occupies the end of the divided 
nerve and is embedded in the cicatricial tissue. In the 
idiopathic form, a clinical feature worth remembering is 
that the tumor may be easily slipped and moved in the 
lateral direction, to and fro, while in the vertical direc- 
tion its position can scarcely be changed. 
While these little tumors are exceedingly painful, they 
are entirely innocent in character. 
io. The Angeiomata.—These are tumors made up 
principally of blood-vessels, and commonly known as 
“claret-cheek,” or “ mother’s marks.” In professional 
parlance such tumors or growths are called nevi, and may 
be divided into two classes—the cutaneous and the sub- 
cutaneous,—according to the volume or character of 
the blood supply, whether from a mesh of capillaries, or 
from larger vessels. 
In the cutaneous or capillary variety the color of the 
“ mark ” is bright-red, while in the subcutaneous variety 
it is a dark purple. 
The growth of these ugly tumors and disfigurements 
is often very rapid, but they involve no danger to life. 
They are liable to either cystic or ulcerative degenera- 
tion ; sometimes they spontaneously disappear. 
n. The Lymphangeiomata.—Our knowledge of 
the pathologic histology of this interesting variety of 
tumors is yet extremely unsatisfactory, and the most 
that can be said at present may be briefly told as fol- 
lows : These tumors are probably cavernous angeiomata 
made up of lymphatic vessels. As to their origin, they THE SARCOMATA. 
151 
may be either congenital or acquired. In the former 
class, the tongue, lips, and genital labia are the parts 
most frequently involved. In the latter class of cases, 
tumors as large as an orange are sometimes met with 
occupying the subcutaneous tissue, especially on the thigh 
and thorax. 
THE SARCOMATA. 
Unlike the group of tumors just described, sarcomata 
do not consist of any of the tissues of the adult body, 
but in structure follow the modified type of some one 
or other of the connective tissues of the embryo. For 
this reason these fleshy tumors are closely related to some 
of the simple growths described in the preceding sec- 
tions of this chapter. 
Following the type of the various connective tissues 
of the body, we have large and small round-cell sarco- 
mata, large and small spindle-cell sarcomata, also the 
giant-cell sarcoma, the mixed-cell sarcoma, the alveolar 
sarcoma, and the melanotic sarcoma. 
Again, we may have such an admixture of cell- 
elements as to warrant the compound names, fibro- 
sarcoma, myxo-sarcoma,glio-sarcoma, chondro-sarcoma, 
osteo-sarcoma, melano-sarcoma, myo-sarcoma, neuro- 
sarcoma, and lympho sarcoma. 
Theoretically, the sarcomata should without exception 
spring from a connective-tissue structure ; but as every 
gland contains a considerable amount of connective 
tissue, a glandular origin of sarcoma is not uncommon. 
Sarcomatous tumors present all degrees of malignancy, 152 
MEDICAL MICROSCOPY. 
and have a peculiar tendency to incorporate connecting 
and adjacent tissues in their growth, thus molding 
themselves upon invading structures. It is a well recog- 
nized fact that the higher the degree of development 
of the tumor the less likely is recurrence after removal. 
And, further, with each recurrence the growth may show 
a more rudimentary structure and greater malignancy. 
This group of tumors may be further described as 
follows:— 
Fig. 39. 
Round-Celled Sarcoma. {Gould.) 
1. The Round-cell Sarcomata.—This variety in- 
cludes both small- and large-cell growths. They are of 
varying density—sometimes being very soft; they are 
of a whitish or pinkish color, seldom encapsuled, and 
while the most rudimentary in structure, they are as a 
rule the most malignant. 
2. The Spindle-cell Sarcomata.—This variety 
must also be subdivided into the small spindle-cell 
and the large spindle-cell sarcomata. The small-cell 
variety may sometimes, by their firm texture and well- THE SARCOMATA. 
153 
defined outline, approach the fibromata. These tumors 
are the “recurrent fibroids” of the surgical pathology 
of long ago. They consist of broad, interlacing bands 
of elongated cells, with but little intercellular structure; 
and after complete removal they show comparatively 
little tendency to recur. 
The large spindle-cell variety is a softer growth than 
the small spindle-cell tumor, and far more malignant in 
character. This form of sarcoma—with its long-tailed, 
large, nucleated cells and but little intercellular sub- 
Fig. 40. 
Giant-Celled Sarcoma. 
{Gould.) 
Small Spindle Celled Sarcoma. 
{Gould.) 
stance,—while rather more common in connection with 
the periosteum, may arise in any fibrous structure. Usu- 
ally, they are but imperfectly encapsuled. These are the 
“ fibro-plastic ” tumors described by the older writers. 
3. The Giant-cell Sarcomata.—This form of 
sarcoma occurs more often in young persons, and in- 
volves by preference the long bones, although a very 
frequent site is the alveolar border of the jaws. To the 154 
MEDICAL MICROSCOPY. 
naked eye a fresh section of such a tumor is pinkish 
in color, but always more or less mottled with darker 
spots from extravasations of blood. Microscopically, 
the so-called giant-cells are irregularly rounded collec- 
tions of granular protoplasm containing numerous round 
or oval nuclei having a well-defined nucleolus. 
In this form of sarcoma, after thorough removal 
recurrence is not probable. In some cases, however, 
the disease is malignant in a high degree. 
Fig. 41. 
Melanotic Alveolar Sarcoma of Skin. (Gould.) 
4. The Mixed-cell Sarcomata.—As the name 
implies, in this form are presented all sorts of cells above 
described. 
5. The Alveolar Sarcomata.—These are rare 
growths, and essentially malignant in character. Micro- 
scopically, they bear a close resemblance to cancer, by 
the coarse stroma which forms the alveolar spaces and 
incloses many or few round nucleated cells. 
6. The Melanotic Sarcomata.—This form is 
really of mixed-cell character, but presents a striking THE CARCINOMATA. 
difference from the ordinary mixed-cell sarcoma in that 
it has a brownish or even blackish appearance from the 
presence of pigment in the cells. Secondary growths 
of like character may take place in any tissue of the 
body. I have known the disease to develop on the 
big toe from a sore corn, and quickly kill the patient. 
These tumors, though often completely encapsuled, 
show a high degree of malignancy. 
7. Glioma.—This is, in fact, but another name for a 
small round-cell sarcoma originating from the neuroglia 
or connective tissue of nerve. 
8. Psammoma.—This rare tumor is found only in 
connection with the membranes of the brain or spinal 
cord. It is made up of flattened cells and is character- 
ized by the peculiarity of containing the so-called 
“ brain-sand. ” The calcified corpora amylacea are held 
together by loose fibrous or highly cellular tissue con- 
taining many vessels. 
9. Teratoma.—This is a congenital tumor. In 
structure it consists of various imperfectly developed 
fetal or mature elements of the body mixed in a hetero- 
geneous manner. Dermoid cysts belong to this group. 
155 
THE CARCINOMATA. 
To Virchow belongs the credit of establishing the 
sharp line of difference between sarcoma and cancer, 
namely:— 
Sarcomata are tumors whose structure fol- 
lows the type of the connective tissues. 156 
MEDICAL MICROSCOPY. 
Cancers are tumors whose structure follows 
the type of the epithelial tissues. And thus, with 
but few exceptions, they may easily be distinguished by 
definite microscopic characters. 
The exceptions referred to have been partially stated 
in the section descriptive of alveolar sarcoma, which so 
closely resembles cancer in microscopic structure as to 
render it very difficult, if not impossible, in some cases 
to make a differential diagnosis without the aid of the 
clinical history. Other examples of difficulty in diagno- 
sis could be furnished ; for instance, it is sometimes hard 
to distinguish between papilloma and epithelioma. 
When, however, there is a well-formed stroma along 
with the epithelial masses, there will be but little diffi- 
culty in making the distinction; but in the study it 
should be remembered that the connective tissue-stroma 
may vary both in quality and quantity, from strong 
fibrous bands and walls around the alveolar spaces, to 
an extremely loose fibro-cellular structure. The alveolar 
spaces containing the epithelial or cancer cells are of all 
shapes and sizes, and differ greatly in their arrangement. 
Cancers never occur elsewhere probably than in con- 
nection with the skin, mucous membranes, and secreting 
glands: in other words, the epidermic or epithelial 
structures—situations where epithelium exists. There 
may be offered some apparent exceptions to this rule, 
and the alleged occurrence of cancer growing into bone 
given in proof; but such extremely rare cases may be 
accounted for by accidental inclusion or graft of em- 
bryonic epithelium at such unnatural sites. THE CARCINOMATA. 
157 
These malignant tumors are not enclosed in capsules, 
as are many of the more benign growths, and hence 
their increase in size by infiltration. 
The glandular type embraces Hard Cancer, or 
Scirrhus, Soft Cancer, or Encephaloid,—includ- 
ing all grades of density between the extremes of hard 
and soft. 
The epithelial and epidermic type includes Cylin- 
Fig. 42. 
Scirrhous Carcinoma. {Gould.) 
Medullary Carcinoma. {Gould.) 
drical Epithelioma and Flat-celled Epithelioma, 
corresponding with the two principal forms of epithe- 
lium. 
Besides these two main divisions, we have Colloid 
Cancer, Melanotic Cancer, and Mucous Cancer. 
Hard Cancer is characterized by a preponderance 
of connective tissue, and occurs most frequently in the 
mamma; but other parts may be the seat of the disease, 
such as the stomach, kidney, ovary, and testicle. As 
this form grows by infiltration and invasion, the organ 158 
MEDICAL MICROSCOPY. 
or part involved may be contracted rather then en- 
larged. 
On section of such a cancer, the cut surface is grayish 
and glistening in appearance, while the tissue is dense 
and elastic, with yellowish lines or markings from fatty 
degeneration of the cells. This form is less malignant 
than the soft variety. 
Soft Cancer is characterized by a very delicate 
stroma and alveoli rather loosely packed with cells, 
usually of smaller size than those found in hard cancer. 
Cancers of this variety may rapidly grow to great size, 
and are malignant in the highest degree. The organs 
and parts most frequently invaded are the ovaries, 
testicles, kidneys, and sometimes the mamma. Owing 
to their soft structure they are freely supplied with 
blood-vessels, and sometimes bleed profusely, forming 
what was formerly called fungus hematodes. 
While it might theoretically be claimed that all 
cancers are epithelial in character, by common usage 
those which invade the skin and mucous membranes, 
and whose malignant tissue proliferates the epithelium 
normally present, are called epithelial, or epithelioma. 
The Cylinder-celled Epithelioma occupies those 
parts covered normally with cylindrical epithelium, and 
has its origin in the mucous membrane, giving the dis- 
eased tissue a glandular appearance. From this fact, 
cylindrical epithelioma may be mistaken for adenoma. 
Indeed, it has been so classed by some writers. 
This form of cancer is chiefly found in the stomach, 
in the intestine—particularly in the rectum,—and very THE CARCINOMATA. 
rarely in the uterus. It grows by infiltrating the mus- 
cular trabeculse. When occupying situations exposed 
to movement or friction, ulceration may take place. 
The Flat-celled Epithelioma occurs on those 
surfaces where there is profuse growth of pavement 
159 
Fig. 43. 
Section of a Cylinder of Epithelial Cells. X 500. «. T'he cylinder itself with 
the characteristic stratification of its cells, a younger and an older pearly 
globule, b. The stroma, very rich in cells at c, and contributing directly to 
the enlargement by apposition of the cylinder. 
epithelium; and so the skin, lips, mouth, pharynx, 
esophagus, larynx, vagina, glans penis, urinary bladder, 
etc., are the sites most frequently invaded. The common 
epithelioma of the lip is a good example of this form of 160 
MEDICAL MICROSCOPY. 
cancer, with its closely packed globes and laminated 
capsules. 
Colloid Cancer is met with chiefly in the stomach, 
intestines, and kidney, rarely in the mamma. Micro- 
scopically, the growth has a gelatinous appearance. So 
distinct are its characters, even to the naked eye, that it 
can scarcely be mistaken for any other form of tumor. 
Melanotic Cancer is nothing more than a soft 
cancer in which pigment granules are deposited in the 
Fig. 44. 
Colloid or Myxomatous Cancer. (Gould.) 
cells and also in some parts of the stroma. It produces 
secondary growths by metastasis, and is exceedingly 
malignant. The parts most frequently invaded are the 
skin and eyeball. 
Mucous Cancer in many particulars resembles col- 
loid cancer. Instead of cell-degeneration into masses 
of colloid material, we have in this form cell-degenera- 
tion into mucoid tissue. The growth presents a gela- 
tinous appearance, and attains its greatest size in the THE CARCINOMATA. 
161 
ovary. But this form of cancer, in any part, is a great 
rarity. 
Mucous and colloid growths arise from the same chem- 
ical principles—the albuminates—and, probably, are 
the results of the conversion of normal tissue-albumin. 
It is mostly in fatty tumors and sarcomas that mucoid 
degeneration takes place; but in some cases it may be 
difficult, if not impossible, to say which of the elements 
—mucus or colloid—is the leading character of the 
growth or degeneration. While the chemical principle 
of mucus is definitely known, colloid is not yet entirely 
understood in its chemical reactions. They may, how- 
ever, be distinguished by their difference in chemical 
reaction : mucin forming a precipitate with acetic acid, 
alcohol, and chromic acid, while colloid is not sensibly 
altered by these reagents. 
The Parasitic Theory of Carcinoma.—This 
idea, suggested by the resemblance of some cancers to 
certain infective diseases, such as tuberculosis, actino- 
mycosis, leprosy, etc., has recently engaged the attention 
of learned pathologists throughout the entire medical 
world. 
Nearly fifty years ago (1847) Virchow—the Great 
Commoner in Pathology—foreshadowed in his plates 
descriptive of his observations on tumors similar bodies 
to those now claimed as parasitic protozoa, but it was 
not until three or four years ago that the important sub- 
ject attracted much attention. In this country the 
question has been most ably discussed by Adler, Coun- 
cilman, Coley, Gibbes, McFarland, Montgomery, Smith, 162 
MEDICAL MICROSCOPY. 
and Van Cott, with a result not in accord with the par- 
asitic theory. But bacteriologic science has given so 
many surprises within the last ten years—so many pos- 
sibilities for the near future—that many of us, though 
unable to give a reason for the faith within us, like the 
patriarchs of old, “look forward to the coming time” 
to make plain all these things. 
There seems to be no doubt that a peculiar body is 
practically always present in some of the cancer cells, 
but its unstable quantity is misleading. It is claimed 
by some observers that the same “parasite” may be 
found also in sarcoma. These inclusions are variously 
rounded, measuring from 2 jj. to 10 /z in diameter, and 
may be single or in eights or tens, and surrounded by 
a capsule. Sometimes the inclusions are seen in the 
cell nucleus, or may occupy any position from the cen- 
ter of the cell-nucleus to the intercellular space. 
Cultivation experiments have not yet succeeded, 
neither has the necessary proof been furnished, that 
cancer may be transmitted by direct inoculation. 
The best results in staining sections of tumors of any 
kind may be accomplished with the Hematoxylin Double 
Stain, as directed on a preceding page. It differentiates 
the so-called cancer “inclusions” in a remarkable 
manner. URINARY EXAMINATIONS. 
163 
XII. URINARY EXAMINATIONS. 
How to Collect a Specimen.—A perfectly clean 
four or six-ounce bottle, well stoppered with a clean 
cork, should be first provided. If the patient or subject 
be a male person, he should be instructed to first wash 
out the urethra by permitting the escape and waste of a 
little urine, and afterward urinate directly into the 
bottle and cork it tightly. If the subject be a female, 
she should first of all cleanse herself of any discharge 
from the vagina by careful washing of the parts, and 
also wash out the urethra by passing a little urine into 
the “chamber,” after which she should collect the re- 
mainder in a sterilized bowl or glass, from which it may 
then be poured into the bottle and immediately corked. 
In no case should the “chamber” vessel be used to 
collect a specimen for microscopic examination, unless 
it be a new one never before used. 
If the specimen cannot be immediately examined, or 
must be kept for several days, the bottle provided for 
this purpose, before being filled, should be thoroughly 
sterilized either in boiling water for five or ten minutes, 
or well baked in the stove-oven. If such necessary care 
be taken and the bottle filled up to the neck as soon as 
possible after collecting the specimen, and tightly 
corked, it will keep without deterioration for many days 
—even weeks. 
To yield a fair average result the specimen should be 
a mixed one, showing both morning and evening urine; 164 
MEDICAL MICROSCOPY. 
but if such mixed urine cannot conveniently be collected, 
two specimens should be taken, one a part of what is 
passed in the morning immediately after rising from 
bed, another a part of what is passed on retiring. 
The urinary excretion expresses the amount of ash or 
waste nitrogenous products eliminated from the body. 
It is perpetually changing in sensible qualities to meet 
the sum of varying impurities, habitual or accidental, 
physiologic and pathologic, of the circulating blood ; 
but the normal average diurnal quantity passed by an 
adult person may safely be set down at from forty to 
fifty ounces. 
Every person understands the direct influence of the 
weather in promoting or restricting the amount of urine 
discharged in twenty-four hours, also the influence of 
large quantities of water taken into the stomach, and 
that from these causes, singly or conjoined, the urine 
may vary both in quantity and quality without being 
inconsistent with good health. If the weather is cold 
and the general surface of the body chilly, there is 
frequent micturition ; if the skin is warm and freely 
perspiring, or the bowels loose, then less urine is dis- 
charged than on the other hand; while if much water 
be drunk, the amount of urine discharged is correspond- 
ingly increased and paler in color. 
Any cause which raises the blood-pressure in the 
vessels, such as by contracting the arterioles, by expos- 
ure to cold, by mental excitement, the influence of 
food, or by the action of drugs, such as digitalis and URINARY EXAMINATIONS. 
165 
scoparius, increases the secretion of urine. On the 
contrary, external warmth, nervous shock, or anything 
else which lessens the blood-pressure in th& glomeruli, 
or raises the pressure in the tubules, diminishes the 
secretion of urine. 
The composition of normal urine per thousand parts 
is as follows :— 
Water, 950.00 
Urea, 26.20 
Creatinin, 0.87 
Sodium and potassium urates, 1.45 
Sodium and potassium hippurates, .... 0.70 
Mucus and coloring matter, 0.35 
Organic. 
Inorganic. 
Sodium biphosphate, 0.40 
Sodium and potassium phosphates, .... 3.35 
Lime and magnesium phosphates, .... 0.83 
Sodium and potassium chlorides, 12.55 
Sodium and potassium sulphates, 3.30 
1000.00 
These proportions, it should be stated, are unstable 
and greatly depend upon age, sex, occupation, diet, 
etc. The formula, however, is sufficiently precise to show 
the fact that water, urea, and the chlorides are the chief 
ingredients of the urine. In other words, that the urine 
is simply a watery solution of urea and organic and 
inorganic salts. 
The color of normal urine is that of wheat-straw, pale 
sherry, or amber; but the color or tint of the vessel 
containing the specimen should not be overlooked in 
describing and comparing the specimen. 
The following very instructive table from Halliburton, 
taken from Prof. Jaksch’s “Clinical Diagnosis,” shows 166 
MEDICAL MICROSCOPY. 
the nature and origin of the chief variations in tint of 
urine: — 
Color. 
Cause of Color. 
Pathologic Condition. 
i. Nearly colorless. 
Dilution or diminution of 
normal pigments. 
Various nervous condi- 
tions, hydruria, diabe- 
tes insipidus, granular 
kidney. 
2. Dark-yellow to 
brown-red. 
Increase of normal or oc- 
currence of pathologic pig- 
ments. 
Acute febrile diseases. 
3. Milky. 
Fat globules. Pus-corpus- 
cles. 
Chyluria. Purulent dis- 
ease in the urinary 
tract. 
4. Orange. 
Excreted drugs, e. g. 
Santonin, chrysophanic 
acid. 
5. Red or reddish. 
Unchanged hemoglobin. 
Hemorrhage or hemo- 
globinuria. 
6. Brown to brown- 
black. 
Pigments in food (logwood, 
madder, bilberries, fuch- 
sin). He mat in. Met- 
hemoglobin. Melanin. 
Hydrochinon. Catechol. 
Small hemorrhages. Met- 
hemoglobinuria. Mela- 
notic sarcoma. Carbol- 
ic acid poisoning. 
7. Greenish-yellow, 
greenis h - 
brown, ap- 
p roac h i ng 
black. 
Bile pigments. 
Jaundice. 
8. Dirty green or 
blue. 
A dark-blue scum on the 
surface with a blue de- 
posit, due to excess of in- 
digo-forming substances. 
Cholera, typhus; seen 
especially when the 
urine is putrefying. 
9 Brown-yellow to 
re d -b r ow n, 
becomes 
blood red on 
addition of 
alkalies. 
Substances introduced into 
the organism, as senna, 
rhubarb, and chelidonium. 
The Specific Gravity of healthy urine varies 
between 1017 and 1023. When for any lengthened URINARY EXAMINATIONS. 
167 
period the mark is found below 1017 or above 1023, it 
is, according to my observation, the sign of a departure 
from the standard of health. In other words, I am 
induced to believe that the standard of perfect health 
carries a urine whose specific gravity is not lower than 
1017 nor higher than 1023. In pathologic conditions 
the range may be widely extended in either direction of 
the scale. 
Perfect accuracy of specific gravity 
can only be attained by testing the 
urine at the exact temperature for 
which the urinometer was construct- 
ed, namely, at 6o° Fahr. At any 
temperatures above this a lower 
specific gravity is indicated, while 
at temperatures below a higher than 
the true one is the result. A new 
urinometer should always be tested 
in distilled water to see if it mark 
correctly 1000 on the scale. 
In using the urinometer care 
should be taken to see that it is clean 
and dry before dropping it into the 
test-tube containing the urinary 
specimen, also that the cylinder 
or tube is large enough to float 
the urinometer without touching the 
side of the cylinder. Any froth or 
bubbles on the surface should be absorbed and removed 
Fig. 45. 
Urinometer. 168 
MEDICAL MICROSCOPY. 
with a slip of blotting paper. When, as often happens, 
the quantity of the specimen brought for examination 
is too small to float the urinometer in its accompanying 
cylinder, it should be exactly measured and as many 
volumes of water added as may be found necessary to 
take the specific gravity. After that, the decimal 
figures should be multiplied by the number of times 
the urine has been diluted, which will give the true 
specific gravity. The transitory variations of specific 
gravity between 1017 and 1023 are of no clinical im- 
portance. 
Reaction.—Fresh normal urine—unless passed im- 
mediately after a hearty meal, when it may be neutral 
or even slightly alkaline—always exhibits a more or less 
acid reaction and reddens litmus, not that it contains a 
free acid, but probably from the presence of the acid 
phosphate of soda. It is diminished by a vegetable 
diet and by alkalies with vegetable acids. 
It has been estimated by Taylor that the acidity of 
the urine during twenty-four hours is equivalent to 
about 14 grains of carbonate of sodium or to 30 
grains of oxalic acid. 
How to Handle the Specimen.—On receipt of 
a specimen for microscopic examination, enough of it 
to measure three or four ounces should be poured into 
a very clean.conical glass, which, after the reaction and 
specific gravity have been taken, should be covered with 
a paper cap to prevent the entrance of dust and dirt of 
all kinds, and then set aside to stand undisturbed for URINARY EXAMINATIONS. 
169 
eight or twelve hours, in order that any morphologic 
constituents present may subside.* Even with the 
greatest care in collecting and subsequent handling, 
foreign matter of some kind is likely to gain entrance, 
such as hairs, fibers of wool, cotton, and flax, minute 
particles of wood, soot, etc. It is, therefore, highly 
important that the observer should first become entirely 
familiar with the appearance of all such common 
objects in the field of the microscope before sitting in 
judgment upon the character of a urinary deposit. (See 
Fig. 6.) 
What has just been said of the great necessity of 
cleanliness in handling a specimen applies with equal 
importance to the pipette, which should be free from 
the possibility of any organized matter carried from a 
previous urinary examination. 
Organized Constituents.—With the pipette, a 
small drop of the deposit is placed at the center of 
the slide and covered with a thin glass. If too much 
fluid has been dropped to hold the cover-glass in place, 
the slide should be put aside for a little while, or until 
sufficient evaporation has taken place to permit it to be 
turned on its edge without slipping the cover. Care 
should be taken that no pressure is made upon the 
cover, lest the object to be studied be unduly com- 
* All reagents and apparatus necessary for urinalysis may be 
obtained from the reliable house of Bullock & Crenshaw, Phila- 
delphia. 170 
MEDICAL MICROSCOPY. 
pressed. It is really necessary sometimes, especially 
when the deposit contains renal casts of large size, to 
give support to the cover-glass to prevent compression. 
Fig. 46. 
Epithelium from the Bladder, 
Ureter, and Pelvis of the 
Kidney. 
Vaginal Epithelium. 
Renal Epithelium, Healtny and Fatty. 
This may be easily accomplished by placing on either 
side of the drop of urine a bit of a fine hair, say a half 
inch in length, which will answer instead of a shallow 
cell, if not better. 
If the sediment is stained before the drop is placed URINARY EXAMINATIONS. 
171 
on the slide, it will greatly facilitate the examination. 
This is readily done by drawing up a pipetteful of the 
sediment and transferring it to a very small vial. A 
drop or two, or as much as may be necessary to give 
appreciable color, of aqueous solution of safranin is 
then added, and the vial shaken for a moment. By 
this simple means all organized matters are more or 
less deeply stained, and the microscopic field can be 
searched more rapidly. 
Epithelial Cells.—If one believe what the books 
say about the different kinds of urinary epithelium, 
there is enough difference in size and shape to distin- 
guish one from the other and say of a certainty, this 
form of flattened, tesselated epithelium comes from the 
renal pelvis ; that form of spheroidal or polyhedral cells 
is from the convoluted tubules; this form of columnar 
epithelium is from the ureter and the greater portion of 
the urethra, or that this flattened or pyriform cell is 
from the bladder. Unfortunately, in practical work no 
such sharp distinctions are present in the microscopic 
field to localize the affection in the urinary tract. 
It is very true that the superficial layers of the mucous 
membrane furnish epithelial cells of polygonal or ellip- 
tical shape with a single large nucleus and granular pro- 
toplasm, while those from the middle and deeper layers 
are of rather oval shape, sometimes running out into 
long protoplasmic tails or processes having also a single 
large nucleus and granular protoplasm—yet all this is 
inferential rather than absolute proof. Quality of 
epithelium without excessive quantity is not of much 172 
value in differential diagnosis. But when both are 
present, there is strong foundation for localizing the 
disease. 
When the kidney cells are found loaded with fat it is 
the indication of fatty degeneration in the renal tissues. 
In women who suffer from leukorrhea, or other disease 
of the vagina and uterine cervix, the urinary deposit is 
usually loaded with large pavement epithelial cells, which 
are sometimes found lying in masses and overlapping 
one another, like the slates on a roof. 
Before closing this section it should be said that the 
most accurate knowledge on the subject does not 
warrant the conclusion that a differential diagnosis can 
be safely founded upon the presence or absence of 
epithelial cells, unaided by other manifestations of the 
urine and the clinical history. 
Mucus.—The meaningless flocculent cloud which 
separates from all urines is well known to every medical 
examiner, and has little if any clinical value. It is 
not a pathologic condition. When, however, mucus 
(nucleo-albumin) in large quantities is passed, it is 
significant of catarrh of some part of the urinary tract 
and then becomes a matter of much importance. 
When this occurs, its presence forms a viscid or gela- 
tinous looking sediment which needs no chemical 
manipulation to determine its character. 
Mucin is always abundantly present in leukemia and 
jaundice, and may be detected chemically by treating 
the urinary specimen with an excess of acetic acid. If 
much mucin is present the urine becomes turbid. 
MEDICAL MICROSCOPY. URINARY EXAMINATIONS. 
173 
Red Blood-corpuscles.—If blood be present in 
moderate quantity the characteristic reddish or smoky 
color cannot well be mistaken by an experienced ob- 
Fig. 47. 
Shrivelled Blood-corpuscles in Urine 
(catarrh of the bladder), with 
numerous lymph-corpuscles and 
crystals of triple phosphate. X 35°. 
(Landois.) 
Colored and (a) Colorless Blood- 
corpuscles of various forms. 
{Landois.) 
Peculiar Changes of the 
Red Blood - corpuscles 
in Renal Hematuria. 
(Landois.) 
Crenated Red Blood-cor- 
puscles in Urine. (Lan- 
dois.) 
server; but when the red corpuscles are but few in 
number, and do not sensibly affect the color, they may 
escape notice without the aid of the microscope. In 
some cases, they may be present in such great abund- 174 
MEDICAL MICROSCOPY. 
ance as to form a layer at the bottom of the glass. The 
shapes and appearances of the red corpuscles may vary 
as much. When they are but few in number, some ot 
them may retain their proper form ; but usually they 
are shrunken in appearance, of pale yellowish color, 
and in general contour look more like washed-out rings 
than normal red blood-cells. In this condition, there 
is ground for suspicion that the blood was effused in 
the kidney, pointing either to acute nephritis—either 
primary or secondary—or tuberculosis. 
When blood is present in large quantity without be- 
ing intimately mixed with the urine, the hemorrhage 
is, in the majority of cases, from the bladder. When 
the blood-cells are so thoroughly mixed that the speci- 
men is deeply tinged, yet no deposit is found after 
several hours’ standing, it may be inferred that the 
source of the hemorrhage is the substance of the kid- 
ney. 
In urine of low specific gravity and ammoniacal 
reaction blood-corpuscles soon lose their color and 
present the appearance of colorless rings ; on the con- 
trary, in urine of normal and even higher than the 
normal specific gravity, with reaction either neutral or 
acid, the cells may retain their color for several days. 
After excluding as factors menstruation, both normal 
and vicarious, purpura, scurvy, mental emotion, etc., a 
careful study should be made to find out if possible the 
exact source of the hemorrhage. To arrive at a just 
conclusion on this point—whether the blood is from 
the substance of the kidney, renal pelvis, ureter, blad- URINARY EXAMINATIONS. 
der, or urethra—other constituents of the urine must be 
carefully examined. While any one of them, per se, may 
be valueless in aiding the differential diagnosis, yet when 
studied all together the inquiry may often result in cor- 
rectly localizing the hemorrhage. 
Profuse and repeated hemorrhage usually accompanies 
malignant disease either of the kidney or bladder. In 
cases of renal abscess, impacted calculus, tuberculous 
infiltration and softening, embolism, or the presence of 
parasites, the hemorrhage is, as a rule, not very profuse. 
175 
Fig. 48. 
Pus-cells in Urine, Unaltered, and Affected by Acetic Acid. (Ral/e.) 
However, I have known profuse hemorrhage to follow 
the passage of a renal calculus. 
Leukocytes—Pus-cells.—It is truly “a dis- 
tinction without a difference,” and most confusing to 
the student, to call these bodies, which are one and the 
same, by different names. When the number found in 
a urinary specimen is but small, they may be called 
leukocytes; when so profuse that they form a thick, 
creamy deposit at the bottom of the collecting-glass, 
they are called pus-cells. 
In every specimen of urine that may be brought for 176 
MEDICAL MICROSCOPY. 
examination a few cells of this character will be found, 
and may be easily recognized with the microscope. 
They are about one-third larger than a red blood- 
corpuscle, and colorless. They are made up of cell- 
wall, granular contents, and nuclei. By the addition 
of a drop of acetic acid, the cell-wall is rendered quite 
transparent and the nuclei more decided. Should 
doubt arise in any case as to their identity, it may be 
quickly dispelled by either the iodo-potassic iodide 
sohition, or Vitali’s guaiacum tincture test. The former 
colors the cells a deep mahogany brown, and the 
epithelium with which they are occasionally blended a 
light yellow color; the latter gives a deep blue tint. 
The only deposit with which pus can be confounded 
is mucus ; but the difference may be discovered in a 
moment by the addition of a little liquor potassa. If it 
be pus, the mass is at once converted into a semi-solid 
viscid precipitate, which when poured out of the test- 
tube holds together like the white of an egg. This is 
Donne’s test. 
While pus-cells may come from an inflammatory con- 
dition in any part of the urinary tract, they are never 
so abundant as in cystitis, unless in case of the sudden 
bursting of an abscess into the tubules or the pelvis of 
the kidney. In cystitis the local symptoms will suffici- 
ently indicate the origin ; while if the suppurating sur- 
face be the urethra no excuse can be offered for a mis- 
take, because the discharge takes place quite independ- 
ently of urination. 
In females, care should be taken to avoid mistake as URINARY EXAMINATIONS. 
177 
to the source of pus in the urine. The precautions given 
on a preceding page concerning the collection of a 
specimen, if observed, will prevent admixture of a 
leukorrheal discharge with the urine. 
Casts.—Nothing in the whole range of urinary 
pathology is of greater clinical importance, perhaps, than 
the subject of casts. For convenient study they may 
be divided into two classes,—the Unorganized and the 
Organized. 
The Unorganized Class includes the “ False- 
Casts,” those formed of crystals and urates and found 
principally in cases of rheumatic or gouty habit, also 
in the urine of infants. Pathologically, these have but 
little significance—at least, no serious importance. 
The Organized Class includes those derived from 
the loops of Henle and the collecting tubes of the 
kidney, namely:— 
1. Blood-casts—consisting of coagulated fibrin 
enclosing blood-corpuscles. These are indicative of 
hemorrhage into the tubules, whether it be due to in- 
tense arterial hyperemia or venous congestion, as in 
acute Bright’s disease. They are soluble in acetic 
acid. 
2. Epithelial Casts—made up of columnar epithe- 
lium or of round cells. They are found in the earlier 
stages of nephritis. The character of the epithelium 
fixes their significance. They are usually abundant at 
first; but at a more advanced period they give place in 
appreciable degree to the hyaline variety. 
3. Hyaline Casts are made up of a translucent, 178 
MEDICAL MICROSCOPY. 
homogeneous, slightly refractive, and often barely 
visible, flexible, proteid material. The large forms 
represent chronicity of the affection, and therefore are 
of graver import. Occasionally, however—but the 
exceptions are very rare,—they are found in acute cases. 
They come both from tubules that have been denuded 
of epithelium, and those that have become dilated from 
contraction of the interstitial substance. 
Small, very pale hyaline casts are sometimes found in 
non-albuminous urine, but this is extremely rare. In 
jaundice, cancer of the liver involving the kidney, 
and in diphtheria they may also be found; so, it is 
seen, their presence has no necessary connection with 
renal disease. They are unaffected by acetic acid. 
4. Waxy Casts are made up of very refractive 
and brittle proteid matter. As a rule, they are longer 
than the others, and when perfect have been aptly com- 
pared to portions of the segmented body of a tapeworm. 
They are so often broken after leaving the kidney that 
in the microscopic field they are seen in short fragments, 
notched, and bearing upon their surface, in some cases, 
white and red blood-corpuscles, fatty globules, arranged 
separately or in confluent masses, either a coating of 
urates or dotted with crystals of various kinds, and 
fungi. They are named from their supposed resem- 
blance to molten beeswax. They are found in all the 
phases of chronic nephritis, in contracted granular as well 
as in amyloid kidneys. In other words, they are not 
characteristic of any particular disease of the kidneys. 
They are unaffected by acetic acid, but frequently URINARY EXAMINATIONS. 
179 
exhibit the amyloid reaction with methyl-violet and 
iodo-potassic-iodid solution. 
5. Granular Casts are variously dark and opaque 
bodies composed of granular material or covered with 
granular cells, and formed by the degeneration of 
hyaline or waxy casts. They differ much in character, 
and no positive diagnosis can be founded upon them 
alone. In color they may be of all shades, from pale- 
yellow to reddish-brown. They are usually seen in 
fragments of various lengths and widths with well- 
defined borders, but the bodies are variously tapered or 
bent. Now and then they are coated with leukocytes 
or pus-cells, fatty globules, and crystals. It has been 
thought probable that they originate in the degeneration 
of blood and epithelial casts. A similar interpretation 
may reasonably be put on casts whose granulation is 
due to fat molecules. This theory has been credited to 
Rindfleisch and Langhans. 
When granular casts in large quantities are found in 
the urine, an inflammatory condition of the kidneys is 
indicated. 
6. Fatty Casts are most frequently found in cases 
of sub-acute and chronic inflammations of the kidney 
of protracted course; hence the unfavorable prognosis 
warranted by their detection in a urinary specimen. 
They are powerfully refracting bodies made up of 
epithelial, hyaline, waxy, or granular casts, filled with 
fatty globules. If the cast be made up of oil globules, 
an advanced stage of fatty degeneration of the kidney 
is fairly indicated. Their presence, however, may be 180 
MEDICAL MICROSCOPY. 
Fig. 49. 
Deposit in “Acid Fermentation” of Urine, a. Fungus; b. amorphous 
sodium urate; c. uric acid; d,. calcium oxalate. 
Fig. so. 
Deposit in Ammoniacal Urine (alkaline fermentation), a. Acid Ammonium 
Urate; b. Ammonio-magnesium Phosphate, c. Bacterium ureae. URINARY EXAMINATIONS. 
181 
misleading in this: that while in the majority of cases 
they are of the gravest import, they may be found dur- 
ing convalescence from acute nephritis. 
From the foregoing descriptions it may safely be said 
that the presence of casts implies disease of the kidney, 
and the advancement or stage of the disease may be 
correctly determined by the association of a particular 
variety of cylinders with certain other bodies in the 
urine. In the microscopic field much difference is 
perceptible in the length and breadth of the cylinders. 
In diameter they range between ioooth of an inch and 
i-5ooth of an inch,—the medium being about the 
i*7ooth of an inch. Their length must be a matter of 
accident, from being broken up after their escape from 
the kidney. They are mostly cylindrical in shape, 
variously tapered, bent, or crooked, and have somewhat 
rounded ends. 
Before Dr. Bright no one had searched the kidneys 
through the urine for the cause of the perilous organic 
malady which has since been distinguished by his name, 
and of which there are often no direct signals excepting 
those furnished by the urine. And this brings the ques- 
tions, incidentally mentioned in the preceding pages, 
concerning one of the most distinctive marks of Bright’s 
disease, namely:— 
“ Does albuminous urine always imply the presence of 
£ Bright’s disease ’? ” 
“ Is Bright’s disease, when present, always accompa- 
nied by albuminous urine ?” 
To both of these important questions the distinguished 182 
MEDICAL MICROSCOPY. 
old medical philosopher of fifty years ago has given an 
emphatic “No.” And then he goes on to give his 
reasons, which are as cogent and as fresh from the 
garner of scientific truth as if written for 1894 instead 
of 1843:— 
“ I believe that some articles of food and some medi- 
cines have the effect, in some persons, of rendering the 
urine for a time albuminous: perhaps it would be more 
correct to say that certain forms of indigestion may 
cause this change. Albumen has also been detected in 
the urine after a blister upon the skin, or under that 
general state of irritation of the surface called eczema 
rubrum, which is produced occasionally by mercury. In 
the crisis of some febrile disorders, in some cases of 
pregnancy, of heart disease, and of delirium tremens, 
and in epidemic cholera, the same phenomenon has 
been observed. Whenever blood, proceeding from any 
part of the long tract of mucous membrane which lines 
the urinary organs, mingles with the urine, that fluid of 
necessity contains albumen, and coagulates if tested by 
heat or by nitric acid. 
“ On the other hand, when the kidney is really affected 
in the way already described, the admixture of albumen 
with the urine is apt to disappear for a while, even sud- 
denly. I have known it to vanish for several hours 
immediately after the effectual application of a hot 
air-bath, and after profuse purging by a full dose of 
elaterium. Sometimes it is absent for a longer period. 
“Another important question, therefore, now arises: 
Finding albumen in the urine, how are we to know URINARY EXAMINATIONS. 
183 
whether it does or does not indicate the presence of 
Bright’s kidney ? 
“We may judge, in part, by frequently testing the 
urine, and noticing whether the albuminous impregna- 
Fig. 51. 
Micrococcus ureae. 
tion be transitory or persistent. If week after week it 
remain steadily present, it is almost surely indicative of 
that renal disease. Almost surely, I say, because it is 
held by M. Rayer, and thought not improbable by Dr. 
Fig. 52. 
Fungi in Urine, e, mould; f, yeast; d, g, micrococci and bacilli; a, b,c, 
uric acid. (Landois.) 
Owen Rees, that uric acid crystals, occurring in the 
urine of gouty persons, may sometimes, by irritating the 
urinary tubules, give rise to enduring albuminuria when 184 
MEDICAL MICROSCOPY. 
there is no degeneration of the kidney. Certain it is 
that in a very few cases albuminuria goes on even for 
years without any serious inconvenience to the patient 
or much visible impairment of his general health. If 
M. Rayer’s pathological doctrine be true at all, it prob- 
ably is true in these cases of abiding albuminuria; and 
in such cases the albumen will readily disappear under 
alkaline treatment, which forms a test of their nature. 
Partly, again, we form our judgment by the absolute 
amount of the albumen in a given measure of urine. If 
the water be deeply charged with that unnatural ingre- 
dient, the presumption is strong that the kidney disease 
is in progress; and when that disease is confirmed, 
another remarkable change is found to have taken place 
in the urine: its specific gravity is very low, and strik- 
ingly in contrast with that of diabetic urine. This is, 
therefore, a very cogent additional diagnostic circum- 
stance.”*— Watson's Practice of Physic. 
* In clinical medicine great surprises never cease, and in no de- 
partment of pathology are they more frequently witnessed than in 
renal affections. In other words, fatal disease of the kidneys may 
be so completely latent that no sign of its presence is given until the 
subject is suddenly struck down. A very remarkable case exactly 
in point occurred a few weeks ago in the practice of Dr. J. F. 
Baldwin, of Columbus, Ohio. The pathologic specimen was sent 
me for microscopic examination, accompanied with the following 
history from Dr. Baldwin:— 
“ The patient was 56 years of age, a well-developed man of active 
business habits, hearty and strong in appearance, who regarded him- 
self as in perfect health. I had been the family physician for sev- 
eral years, but he had never needed my professional service. He URINARY EXAMINATIONS. 
185 
Urea is by far the most abundant as well as the most 
important of the organic constituents of urine, the 
quantity passed per 
diem by a healthy man 
of average weight being 
512.4 grains. It con- 
stitutes 70 or 80 per 
cent, of the entire nitro- 
genous waste excreted 
by the kidneys. 
A ready method of 
estimating approxima- 
tively the quantity of 
urea present in urine, 
when in excess of the 
normal, is by the addi- 
tion of an equal part of 
nitric acid. The test- 
tube containing the 
urine and the acid is 
set in cold water, after 
which—if urea be in excess—the hexagonal crystals of 
Graduated Ureameter. 
was taken sick on Wednesday. I saw him on Thursday, and found 
him with a pulse of 112, strong and hard; temperature 1020. He 
was suffering with an acute bronchitis, and complained of much 
muscular aching and soreness, with great weakness. Owing to his 
age and general condition I at once obtained a bottle of his urine for 
examination. The reaction was acid, and s. g. 1.021 ; no trace of 
albumin. 
“ The next day, Friday, he was more comfortable in every re- 186 
MEDICAL MICROSCOPY. 
nitrate of urea soon make their appearance. Or, another 
simple method, and a very interesting one, too, is the 
following: take an inch of a small thread of cotton, dip 
one end into the urine, and place it with its hanging 
spect; and the next day, also. He had no headache, and made no 
other complaint than of muscular weakness. Owing to this contin- 
ued complaint of muscular weakness, I still suspected a kidney com- 
plication, and at this visit secured a second urinary specimen. This 
had a sp. gr. of 1.025—still, no trace of albumin. A slide was 
examined for casts with negative result. 
“ Sunday, the fifth day of his illness, he was inclined to be 
drowsy and at times a little delirious. When pressed by the family 
for a statement as to the exact nature of his condition, I said: * I 
can regard his case as only one of grippe, with bronchitis; but if 
the urinary examination had shown any evidence of Bright’s dis- 
ease, that would fully explain his entire condition.’ He had taken 
a dose of physic on Wednesday which operated freely. At no time 
was there vomiting; appetite unimpaired. 
“Monday, the sixth day, he was more stupid, and about midnight 
could not be roused. He then had a temperature of 105°, pulse 
160, contracted pupils, and was bathed in profuse perspiration; in 
other words, he presented a very perfect picture of uremic coma. 
He died at 5 A. M., Tuesday. 
“A specimen of urine passed on Monday evening had a sp. gr. 
of 1.035, and showed a mere trace of albumin. 
“Post-mortem, held Wednesday morning, revealed kidneys larger 
than normal and very pale. There was marked thickening of the left 
ventricle, but no evidence of atheroma. Certainly there was not the 
least evidence, by naked eye appearances, of ulcerative endocarditis. ” 
The structure of the kidney was found literally riddled with 
hemorrhagic infarctions and abscesses, from the size of a millet seed 
to a large bean, with marked changes along the lines of the capilla- 
ries and around the Malpighian bodies. URINARY EXAMINATIONS. 
187 
drop at the center of a slide, then cover with a thin 
glass, leaving half of the thread free. Upon this free 
Fig. 54. {After LanJois.) 
Hyaline Casts, a, b, with 
Leukocytes; c, with 
Renal Epithelium. 
Epithelial Casts. 
Blood Cast. 
a. Leukocyte Cast. b. Acid Sodic 
Urate in Cylinders. 
Finely Granular 
Casts. 
Coarsely Granular 
Casts. 188 
MEDICAL MICROSCOPY. 
end let fall a minute drop of nitric acid, and place the 
slide under the microscope. The formation of the plate 
crystals will afford an interesting field. 
In Bright’s disease the urine is usually diminished. 
Spermatozoids.—These bodies when once seen can 
never afterward be mistaken for anything else. They 
Fig. 55. 
Normal Semen, a. Spermatozoa, b. Seminal cells, c. Epithelium, d. 
Seminal granules. 
are found in the urine of both males and females when 
passed after coitus, and may be of value in aiding the 
diagnosis, as in “ Case i,” recorded in the introductory 
chapter. Their discovery may also aid in establishing 
truth in a medico-legal inquiry, and thus be the means 
of saving the innocent or punishing the guilty. URINARY EXAMINATIONS. 
189 
Tubercle Bacillus.—The recognition of tubercle 
bacilli in the urine is a matter of the very highest 
pathologic importance. A case has just passed from 
under my observation in which the discovery of this 
microorganism in the urine was the means of informing 
the attending physician of the true nature of his patient’s 
ailment. 
The method of demonstrating the tubercle bacillus 
and other microorganisms in urine is the same as that 
already fully described for a cover-glass preparation of 
the sputum. 
Other Organisms.—Healthy urine after standing 
and undergoing the alkaline fermentation contains 
swarms of bacteria exhibiting their characteristic move- 
ments. Various forms of torula are found in diabetic 
urine—the Torula cerevisice and Penicillium glaucum; 
also certain entozoa, the most important of which is the 
Bilharzia hematobia. Besides these, hooklets and frag- 
ments of echinococci from a ruptured hydatid cyst may 
occasionally be detected in the urine. 
Urates.—If after standing and cooling, the urine, 
which was clear when passed, becomes muddy in appear- 
ance and soon shows more or less sediment of a pinkish, 
brick-dust, or deep-red color, it may be taken that the 
deposit consists of urates. If it dissolve by heat and 
the urine becomes again as clear as when passed from 
the bladder, then there can be no doubt of the presence 
of urates. 
Brick-dust colored urates are inseparable from an acid 190 
MEDICAL MICROSCOPY. 
urine. The murexid test for uric acid is equally applic- 
able to urates. This condition of the secretion is com- 
monly seen in concentrated urines. 
If the urine be acid and contain a white flocculent 
deposit, the sediment probably consists of pus, or is 
made up of alkaline urates called the “ Pale White 
Urates.” If such be its composition, the deposit is 
insoluble by heating, but clears by the addition of acetic 
acid. As a rule, the sediment is phosphatic if the urine 
when passed is of alkaline reaction. 
An abundant deposit of urates accompanies the febrile 
state, gout, rheumatism, disease of the liver and kidneys, 
and some forms of dyspepsia. It is frequently seen in 
healthy subjects after great muscular exercise and pro- 
fuse perspiration. 
On the other hand, alkaline urine and phosphatic 
sediments accompany nervous exhaustion, “ nervous 
dyspepsia,” and the vigils of care-worn business men. 
It is also common in the nursery, during the period of 
dentition, in the form of “white phosphates,” and, 
likewise, in nursing women of run-down habit. 
Uric Acid.—Some specimens of urine are so sur- 
charged with uric acid that cayenne-pepper-like grains 
adhere to the sides of the bottle and are also thrown 
down in the deposit. In microscopic appearance the 
crystals vary greatly both in size and shape. Some of 
them are six-sided plates, others four-sided rhombs, 
others oval, others barrel-shape, others comb or whet- 
stone shape, and still others in rosettes,—appearing URINARY EXAMINATIONS. 
191 
either singly or in heaps. In color, they are brownish 
yellow (due to uroerythrin), owing to the affinity of the 
urinary pigment for uric acid and its salts. 
If free uric acid shows itself in freshly passed urine, a 
deposit in the bladder or in the pelvis of the kidney 
Fig. 56. 
i. Rhombic Plates; 2. Whetstone Forms; 3. Quadrate Forms; 4,5. Prolonged 
into Points, 6,8. Rosettes; 7. Pointed Bundles; 9. Barrel Forms precipi- 
tated by adding Hydrochloric Acid to Urine. (Landois.) 
may be taking place and lay the foundation of a cal- 
culus. 
After standing for some time, if the vessel containing 
it was not properly sterilized, the acidity of the specimen MEDICAL MICROSCOPY. 
192 
becomes fainter and fainter until at length it is neutral; 
this quickly gives place to alkalinity, and then a putres- 
cent ammoniacal odor is given off. 
Oxalate of Lime.—Crystals of this substance are 
exceedingly common in both acid and alkaline urine. 
They occur as strongly refracting octahedra, or four- 
sided pyramids of various sizes, also as colorless dumb- 
Fig. 57. 
Oxalate of Lime. 
bells, and when transient, have no clinical significance. 
They have been found in healthy urine, especially after 
the ingestion of fruits and vegetables containing a large 
proportion of oxalic acid, such as asparagus, rhubarb, 
tomatoes, etc. They are soluble in hydrochloric acid, 
but are unaffected by acetic acid. 
The characteristic form of the crystals scarcely admits URINARY EXAMINATIONS. 
193 
of confusion with other forms. The only crystals with 
which it is possible to confound them are the triple 
phosphates, which are larger. Besides other differences, 
acetic acid dissolves the triple phosphates but has no 
effect on the oxalate of lime. 
Persons whose urine is constantly marked by the 
presence of crystals of the oxalate of lime are moody 
and more or less hypochondriacal. This condition has 
been termed the “oxalic acid diathesis,” and the 
particular disease, oxaluria, has a long train of trouble- 
some symptoms ; but the chief significance belonging 
to the deposit is the possible formation of the mulberry 
calculus. 
Triple Phosphates.—The precipitates of the 
earthy phosphates of lime and magnesium so commonly 
encountered in urinary examinations present themselves 
in three forms, namely— 
The amorphous lime phosphate. 
The crystalline lime phosphate. 
The ammonio-magnesium phosphate. 
The amorphous lime phosphate is insoluble in water, 
but soluble in acetic acid. Urine containing it has 
a milky appearance, and on standing soon deposits 
a grayish-white sediment, which the beginner may mis- 
take for pus. Donne’s test will show the distinction. 
Heat precipitates the lime salts in flakes resembling 
albumin, and in this way I have known mistakes to be 
made. 
This character of urine is usually the sign of ner- 
vous exhaustion from any cause, especially from pro- 194 
longed 'mental worry and anxiety, loss of sleep, and 
excessive venery. 
The crystalline lime phosphate presents itself in 
colorless needle-like crystals, called stellar phosphates, 
which arrange themselves in radiating bundles. 
The ammonio-magnesium phosphate is the result of 
MEDICAL MICROSCOPY. 
Fig. 58. 
Forms of Crystals of the Ammonio-magnesium Phosphate. (Tyson.) 
alkaline fermentation and the decomposition of urea. 
These crystals of basic magnesium phosphate are very 
characteristic and are easily recognized by their many- 
sided angles, size, beveled edges, large, strongly re- 
fracting, elongated rhombic tablets, and coffin-lid 
crystals. Although usually found in alkaline urine, URINARY EXAMINATIONS. 
195 
still they are sometimes met with in feebly acid urine. 
A greasy-looking scum frequently forms on such urine. 
In some examples, beautiful stellar and feathery crystals 
are shown, as in the accompanying illustrations. 
Clinically, their presence is of but little consequence. 
Stellate and Feathery Crystals of Triple Phosphate. (Tyson.) 
Urates of Ammonium and Sodium.—Crystals 
of urate of ammonium are easily recognized by their 
spherical form, dark color, and radiating spicules, 
known as hedgehog crystals. Crystals of sodium urate 
are met with in the chalky concretions common in the 
bodies of gouty persons, and also in the urine, where 196 
MEDICAL MICROSCOPY. 
they assume a globular form, with irregular projections 
over the surface. The pathologic significance of the 
crystalline urates is much more important than that 
which attaches to the amorphous deposits. 
Fig. 6o. 
Acid Ammonium Urate. (Lahdois.) 
Sulphate of Calcium.—This substance takes the 
form of long, colorless tables with abrupt ends, also in 
needle-shape. They are rarely met with, and are neither 
soluble in ammonia nor acids. They have no clinical 
importance. 
Chylous Urine.—Chyluria has been a puzzling 
subject to pathologists. The urine has a milky ap- 
pearance, and contains more or less blood, fibrin, and 
albumin. Spontaneous coagulation has been known to 
take place in the bladder and block up the urethra. 
Renal casts have been searched for in vain, and it is 
believed by some authorities (Roberts among them) 
that the chyle and lymph are discharged directly into 
the urinary passage from the lymphatic vessels. It is, 
fortunately, a rare affection. In all of my experience I 
have known but one case, a passenger conductor on the URINARY EXAMINATIONS. 
197 
B. & O. R. R., who, though discharging a large quan- 
tity of chylous urine every day, was in the enjoyment 
of ordinary good health. His run was from Baltimore 
to Wheeling—379 miles—and when on duty the quan- 
tity was always much increased. After sound sleep, the 
flow was, as a rule, sensibly diminished. 
Fig. 6i. 
a. Crystals of Cystin. b. Oxalate of Lime. c. Hour-glass Forms of b 
Cystin.—Urine containing cystin is usually feebly 
acid, of a yellowish-green color, and of peculiar odor, 
which has been compared to sweet briar, has an oily 
appearance, and decomposition takes place very soon 
after being voided. The deposit is a light rose-colored 198 
MEDICAL MICROSCOPY. 
powder. The crystals are symmetrical six-sided plates 
of various sizes, and either appear in overlapping masses, 
or are contiguous to one another. The liver is supposed 
to be the source of this substance. Its most important 
clinical significance is its liability to form a calculus. 
But such cases are, fortunately, very rare, as shown in the 
interesting paper read by Prof. James Tyson, of Phila- 
delphia, before the Association of American Physicians, 
Eighth Annual Meeting, 1893. 
The crystals are insoluble in acids, but readily soluble 
in ammonia. 
Fig. 62 
Hippuric Acid. (Landois.) 
Hippuric Acid.—Hippuric acid may be encoun- 
tered as a crystalline deposit during the acid fermen- 
tation of the urine. It may occur in considerable 
quantity after the exhibition of benzoic acid, or the 
ingestion of green gages, whortleberries, and cranber- 
ries, but it has no clinical significance. 
Xanthin.—This occurs in the form of whetstone URINARY EXAMINATIONS. 
199 
crystals, which are insoluble in acetic acid and soluble 
in ammonia. 
Tyrosin and Leucin.—Tyrosin crystals have the 
form of beautiful sheaves of very fine needles, while 
leucin, though mainly held in solution, presents itself 
a. a. Leucin Balls, b. b. Tyrosin Sheaves, c. Double Balls of Ammonium 
Urate. 
in the form of variously sized spheres. Tyrosin and 
leucin have been found in the urine of persons 
poisoned with phosphorus, also in cases of acute 
yellow atrophy of the liver, yellow fever, and several 
other infectious diseases. 200 
MEDICAL MICROSCOPY. 
Cholesterin.—Crystals of cholesterin are very rarely 
seen in the urinary sediment. 
Fig. 64. 
Cholesterin. (Landois.) 
Summary.—With certain precautions the following 
summary maybe useful to remember :— 
1. Heat precipitates albumin and phosphates, and 
dissolves urates. 
2. Nitric acid precipitates albumin, urea, and uric 
acid and dissolves phosphates. 
3. Liq. potassa precipitates phosphates, dissolves 
albumin, and glutinizes pus. 
4. Acetic acid precipitates cystin and dissolves albu- 
min. 
5. Nitrate of silver precipitates chlorides. 
6. Barium chloride precipitates sulphates. THE BLOOD. 
201 
XIII. THE BLOOD. 
The volume of crimson liquid, whose office is to 
carry nutriment and oxygen to the tissues, is about eight 
per cent, of the total weight of the body. It is of 
alkaline reaction, has a specific gravity of from 1.059 to 
1.062, and composed of three classes of cells, namely :— 
(a) Biconcave circular yellowish discs. 
(&) White blood-corpuscles (leucocytes) 
(<r) Blood-tablets, or platelets. 
The red blood-cells have a diameter of 1-3.200 (7.5 
fi) of an inch ; the white cells 1-2.500 (10 fi) of an 
inch in diameter; and the blood-platelets, the third 
class of formed elements, about one-third the diameter 
of a red blood-corpuscle. These are, of course, average 
measurements. 
The normal number of red corpuscles in the blood is 
5,000,000 in a man and 4,500,000 in a woman to the 
cubic millimeter. 
In normal blood, the proportion of white to red cor- 
puscles is about 1 to 300. The white corpuscles (leuko- 
cytes) are granular spheroidal bodies possessing ameboid 
movements, and perform the office of phagocytes in 
taking up bacteria in the circulating blood. One or 
more nuclei are present, but it may require a little 
weak acetic acid to be run under the cover-glass to 
detect them. To demonstrate the blood-platelets, re- 
quires the addition of some fixing medium, preferably 202 
MEDICAL MICROSCOPY. 
Hayem’s solution, as represented in the following 
formula:— 
Chlorid of sodium, I grm. 
Sulphate sodium, 5 grm. 
Corrosive sublimate, 0.2 grm. 
Distilled water, 200 grm. 
Fig. 65 
Blood-corpuscles, and Blood-plates, from Normal Human Blood, a. Red 
Blood-corpuscles, b. Colorless Corpuscles, c. Blood-plates. (Jaksch.) 
Wash the end of the finger in clean water, then prick 
it with a needle and let the first drops of blood be al- 
lowed to flow off. After this is done, take a clean slide 
and at its center touch the hanging drop without bring- 
ing the glass in contact with the finger, then add a drop 
or two of the solution to the blood, and, as quickly as THE BLOOD. 
possible, cover the sample with a thin glass. The plates 
will then be shown. To prevent evaporation, the cover 
glass should be run around with oil or balsam. 
Arterial blood derives its scarlet color from the pre- 
sence of oxygen which controls the amount of hemo- 
globin contained in the red corpuscles. In other words, 
when the blood is rich in oxygen the coloring matter is 
increased. In the capillaries the blood gives up oxygen 
to the tissues and becomes the dark venous blood. Any- 
thing which interferes with the absorption of oxygen in 
the lungs and the giving off of carbonic anhydride pro- 
duces instant and marked change in the chemical 
quality of the circulating blood. But it should be re- 
membered that in certain morbid states arterial blood 
may assume even a brighter than the normal color, e.g., 
in carbonic oxid poisoning, in which condition it 
becomes of a bright cherry-red, alike in the arteries and 
veins. 
In venous blood there is less oxygen, and hence the 
color is darker, or bluish red. The blood taken from 
the tip of the finger for microscopic examination is 
usually of venous character. 
The chief diseases of the blood are anemia, pernicious 
anemia, and chlorosis, in which the red corpuscles are 
deficient; melanemia, in which pigment-masses are 
formed, with destruction of the red corpuscles ; leu- 
kocytosis, in which the white corpuscles are moderately 
increased, and leukocythemia, in which they are inor- 
dinately increased; and conditions in which morbid 
substances are present, such as the poisons of uremia, 
203 204 
MEDICAL MICROSCOPY. 
cholemia, etc., or microorganisms and 
their products, such as pyemia, septi- 
cemia, etc., “blood poisoning” in the 
ordinary sense. 
Various costly instruments have been 
devised for counting the precise number 
of the red and white blood-corpuscles 
or the relative quantity of hemoglobin 
present in a given specimen, but the 
principle on which they are constructed 
is the same. Probably the simplest and 
best instrument is the Thoma and Zeiss 
apparatus for counting blood-corpuscles. 
It consists of a capillary tube (Fig. 66) 
of glass, about io centimeters long, ex- 
panding in its upper third to a bulb, in 
which lies a small glass ball. The lower 
end of the tube is furnished with a 
scale, graduated in parts, number o.i, 
0.5, 1, up to 101. With this instru- 
ment is used a counting-chamber, in- 
vented by Abbe and Zeiss. This is a 
glass receptacle cemented upon a glass 
slide (Fig. 67); it is exactly 0.1 mm. 
in depth, and its floor is marked out 
into microscopic squares. The space 
overlying each square, 1-4000 cubic 
mm., and the squares, are portioned 
out in groups of 16 by plainer lines 
(Fig. 67). 
Capillary Tube. Thoma-Zeiss Apparatus for Counting the Blood Corpuscles. (Jaksch.) 
Fig. 66. THE BLOOD. 
205 
“ Application of the Process.—A puncture is 
made in the tip of the finger, with the precautions 
already indicated, and the blood then sucked into the 
pipette from the exuding drop to division 1. The point 
of the pipette is wiped, and a three per cent, solution 
of common salt sucked in until the fluid has risen to 
the point marked 101. The contents of the tube are 
then thoroughly mixed, and the column of fluid in the 
capillary tube is removed by blowing into the tube, as 
Fig. 67. 
Counting Slide. 
the blood does not mix with the solution of common 
salt. The neglect of this precaution, therefore, would 
vitiate the experiment. 
“ The hollow cell of the slide is next filled with the 
mixed blood-and-salt-solution, care being taken to 
guard against the admission of air bubbles, and the cover- 
glass is accurately adjusted in such a manner that New- 
ton’s color-rings are produced. The preparation is left 
to stand a few minutes, so as to allow of an intimate 
admixture of its parts, after which it is placed under 
the microscope, and examined with a power of 30-70 206 
diameters, when it will be seen whether any air-bubbles 
or foreign bodies are present in it, and whether the cor- 
puscles are pretty evenly distributed through the fluid. 
The latter are then counted under a high power. In 
doing this, the number present in sixteen squares is 
counted, and from this the average is estimated. The 
greater the number of the squares taken, the more accu- 
rate will be the result attained.” 
After use, the capillary tube must be thoroughly 
cleansed, first with water, then with alcohol, and lastly 
with ether. 
To estimate the number of white corpuscles in a 
specimen of blood it should be diluted with water con- 
taining one-third per cent, glacial acetic acid in the 
proportion of i : io. In this way the red corpuscles 
are destroyed, and the white alone remain in the field 
of vision. In mixing the fluids, a mixing glass speci- 
ally devised by Zeiss for the purpose may be used. The 
process may be carried out thus :— 
“By means of a pipette of i c.c. contents, and accu- 
rately graduated in o. i c.c. units, 0.9 c.c. of the acetic 
acid solution is measured out into a watch glass; with 
another pipette holding exactly 0.1 c.c., the blood is 
added to this and the two well mixed. A drop of the 
mixture is placed within the counting-chamber of the 
cytometer prepared as above ; and now, since the num- 
ber of corpuscles is relatively fewer, the entire field, and 
not the marked-out divisions, is taken as the basis of 
calculation, greater accuracy being so obtained. To 
the same end, a lower power will be used, so as just to 
MEDICAL MICROSCOPY. THE BLOOD. 
207 
bring the marks in the floor of the chamber clearly into 
view. Before beginning to count the corpuscles, how- 
ever, it will be well to focus with the fine adjustment of 
the microscope, and to make sure that the cells have 
all settled. 
“ When there is a very great increase in the number 
of leukocytes, as in leukemia, their number can be 
estimated in the same manner as that of the red cor- 
puscles, and the relative proportion of the two can be 
determined at the same time with sufficient accuracy, 
if only an adequate number of squares is taken into 
account. Great assistance in such experiments may be 
derived from the use of a three per cent, salt solution 
colored with gentian violet, in which the leukocytes are 
stained, and become readily discernible from the red 
blood-corpuscles, which are usually somewhat paler than 
normal. For the same purpose Toison employs a stain- 
ing fluid of the following composition :— 
“ Distilled water, 160 c.c 
Glycerin, 30 c.c. 
Sulphate of soda 8 grm. 
Chlorate of sodium, 1 grm. 
Methyl-violet, 0.025 grm.” 
(Jaksch's Clin. Diagnosis.) 
Test for Hemin Crystals.—The supposed blood- 
coloring matter must be dried, finely powdered, and 
placed upon a slide. A crystal of common salt is 
then added to it and over all a cover-glass is laid. A 
few drops of glacial acetic acid are then allowed to flow 208 
MEDICAL MICROSCOPY. 
under the thin glass. The slide is then placed on a 
heating table, and the temperature run up to 200° F., 
Fig. 68, 
Hemin or Teichmann's Crystals. (Landois.) 
or a little below the boiling point, for a moment. If the 
suspected specimen contains blood-coloring matter the 
microscopic field will show hemin crystals. 
Fig. 69. 
Leuksemic Blood. (Landois.) 
Poikilocytosis.—By this term is indicated a re- 
markable condition of the red corpuscles. It was first 
noticed in pernicious anemia, and by some authorities THE BLOOD. 
209 
is regarded as pathognomonic of that affection. The 
red corpuscles are of all sizes and shapes, so that the 
Fig. 70. 
condition is easily recognized with the microscope. 
Such condition, however, is not confined to any one 
Blood in Poikilocytosis 
Fig. 71. 
Filaria sanguinis hominis. Lewis. (From Leuckart, after Lewis.) 
disease, and it is possible that the poikilocytes do not 
exist in the circulating blood. 210 
MEDICAL MICROSCOPY. 
Animal Parasites.—These are the filaria sanguinis 
hominis(Fig. 71) and the bilharzia hematobia (Fig. 72). 
Fig. 72. 
Bilharzia hematobia, Cobbold, Male and Female. The latter in the 
Canalis gynaecophorus of the former. (After Leuckart.) 
These parasites are also found in the urine, but instances 
either in the blood or urine are extremely rare in the 
United States. 
It has now become a matter of routine in clinical 
medicine to examine the sputum microscopically in all 
cases in which cough and expectoration are present. 
In a preceding chapter, full instructions have been 
given for the most important examination that can be 
made of the sputum, namely, that for the demonstra- 
tion of the tubercle bacillus; and as the technique is 
XIV. THE SPUTUM. THE SPUTUM. 
211 
the same for all other cover-glass preparations, the reader 
is referred to page 116. 
Diphtheritic Membrane and Other Exudates. 
—Examination of such specimens may be made by 
spreading them out on the slide, and staining with 
aqueous solution of either methyl-blue or fuchsin before 
applying the cover-glass. But permanent mounts are 
so easily made of such material and the demonstration 
Fig. 73. 
Microscopic Characters of Mixed Saliva and the Buccal Secretion, a. Epithe- 
lial cells, b. Salivary corpuscles, c. Fat droplets, d. Leukocytes, e. Spiro- 
chaeta buccalis. f. Comma bacillus of the mouth, g. Leptothrix buccalis. h. 
i. k. Different forms of fungi. (Landois.) 
of the microorganisms so much more satisfactory when 
thus treated, that all such specimens should be cast in 
paraffin and given the highest differential staining. 
Crystalline Forms.—Various crystals are found in 
the expectoration, and among them none more common 
than the Charcot-Leyden crystals expelled by asthmatic 
patients. They are diamond-shaped bodies, long and 
pointed (Fig. 74). These crystals are soluble in 212 
MEDICAL MICROSCOPY. 
alkalies, mineral acids, and warm water ; but insoluble 
in alcohol, ether, or cold water. Although their chem- 
ical composition is not yet definitely understood, it is 
supposed that they are the result of decomposition of 
organic matter in the bronchioles, probably Cursch- 
mann’s “spirals.” While frequently found in large 
Fig. 74. 
Charcot-Leyden Crystals. (Landois.) 
numbers in asthma, their presence is not positively 
diagnostic of that disease. 
In phthisis, chronic pneumonia, pulmonary abscess, 
etc., cholesterin crystals (Fig. 64) are frequently met 
with. 
Crystals of the fatty acids, often seen in rosette 
form, are sometimes found in connection with bronchi- 
ectasis and phthisis, also in gangrene of the lungs. FECAL DISCHARGES. 
213 
They consist chiefly of palmitic and stearic &cids, and 
have but little diagnostic value. 
Besides the forms just described, leucin and tyrosin, 
oxalate of lime, triple phosphates, and concretions of 
lime salts are found among the matters coughed up. 
The feces have been aptly compared to “ a garden 
full of weeds.” The quantity passed by a healthy man 
in twenty-four hours averages from five to seven ounces, 
and is composed of a great variety of refuse-substances 
derived from the food, viz., vegetable cells, muscle 
fibers, elastic fibers, connective tissue, fat, starch gran- 
ules, coagulated proteids (especially in the stools of 
young children fed on milk and in persons suffering 
from diarrhea); red blood-cells and leukocytes, epithe- 
lium ; non-pathogenic and pathogenic vegetable para- 
sites ; animal parasites—Protozoa and Vermes. In the 
last named class (Platodes) are the following tape- 
worms :— 
Taenia saginata or “ beef” tapeworm. 
Taenia solium, sometimes called the “ pork ” 
tapeworm. 
Taenia nana.—This is very common in Italy and 
Sicily, particularly attacking children and young 
persons. 
Taenia flavopunctata, bearing a close resemblance 
to T. nana, but of greater length. 
Taenia cucumerina (elliptica).—This parasite is 
XV. FECAL DISCHARGES. 214 
MEDICAL MICROSCOPY. 
common in human beings, being acquired from dogs, 
and especially infests children. 
Bothriocephalus latus.—This worm has recently 
been much studied and acquired clinical importance 
from having been found frequently in cases of pernicious 
anemia. 
In the Class Annelides, or round worms, we have 
Fig. 75. 
Cephalic End of Taenia solium. 
Linne. (After Leuckart.) 
Measly Pork. (After 
Leuckart!) 
the Ascaris lumbricoides, common to all climates, and 
in persons of all ages. It also infests cattle and sheep, 
but is without special clinical import, notwithstanding 
the general impression among unprofessional people that 
its presence causes belly-pains and cramps and spasms 
in children. 
Oxyuris vermicularis, or common thread-worm. FECAL DISCHARGES. 
215 
—The presence of this worm gives uncomfortable itch- 
ing and other disagreeable sensations about the anus. 
Dochmius (Anchylostoma) duodenale.—This 
Fig. 76. 
Taenia saginata. Gceze. (After Leuckart.) 
worm was long since reported as occurring both in 
America and the West Indies; (Louisiana, Chalbert, 
1830; Duncan, 1840; Alabama and Georgia, Lyell, 216 
MEDICAL MICROSCOPY. 
1849). No recent observations seem to have been 
made, and a careful search for the eggs in the feces of 
patients presenting symptoms of pernicious anemia 
without apparently sufficient cause should be carried 
out. 
Trichocephalus dispar (Whip Worm).—This 
Fig. 77. 
Trichina spiralis. Owen. a. Encapsulated with connective tissue covering 
(in situ): b. Calcified. (After Leuckart.) 
worm is supposed to have causal relation with the beri- 
beri disease which is endemic in Sumatra.* 
Trichina spiralis.—This pest occurs in two dif- 
ferent forms in the human body, according to its partic- 
ular site in the muscular tissue or in the intestine. 
* Report of the Ceylon Commission, 1887. MICROORGANISMS OF THE SKIN. 
217 
XVI. MUCOUS CYLINDERS AND CRYS- 
TALLINE BODIES. 
The cases in which mucous formations and crystal- 
line bodies are found in the feces are exceedingly 
common. The passage of casts or cylinders is usually 
accompanied with abdominal pains and tenesmus. The 
presence of such a specimen by stool implies catarrhal 
inflammation of some part of the intestinal tract, 
usually the large intestine. 
All of the crystalline forms that have been named 
as occurring in sputum, including hematoidin crystals, 
are more or less common also in the feces. 
The microscopic appearances of the feces may be 
studied by pressing a minute particle between a cover- 
glass and the slide. If to the specimen is added a drop 
of aqueous solution of methyl-blue, the examination 
will be much more satisfactory. 
XVII. MICROORGANISMS OF THE 
SKIN. 
Most of the microorganisms having their habitat 
on the skin in the cutaneous secretions are sapro- 
phytes, but pathogenic varieties are also here found. 
The space beneath the finger nails affords a rich source 
for various germs. Such specimens, after being freed 
from fat by means of alcohol and ether, may be stained 
with borax and methyl-blue (No. 7) for a few minutes; 
then for half a minute or a minute in a one-half to 218 
MEDICAL MICROSCOPY. 
one per cent, aqueous solution of resorcin, and lastly 
for a few minutes with alcohol. 
XVIII. MICROORGANISMS IN FOODS 
AND DRINKS. 
Many food-stuffs, owing to exposure to various sources 
of contamination, particularly milk and meat, become 
sources of infection. Of all the food-carriers of bacteria, 
milk is the most dangerous. It is the most active 
vehicle for the dissemination of tuberculosis and enteric 
or typhoid fever, except dried sputum, which science has 
discovered and exposed. 
Besides contamination either during milking or in the 
subsequent handling, various pathogenic microorganisms 
may be imparted to milk from the animal furnishing it. 
To examine a specimen of milk, a drop should be 
mixed with another of distilled water upon the slide. 
After drying in the air of the room, the film should be 
fixed by heat, but not scorched. It is now ready to be 
stained with a few drops of alcoholic solution of methyl- 
blue with an equal quantity of chloroform. After the 
evaporation of the chloroform, any excess of the blue 
is washed away in alcohol, and the mount finished in 
the usual way. 
THE BACTERIOLOGIC EXAMINATION OF 
DRINKING-WATER. 
In The Medical News for June 14, 1890, p. 641, and 
in the American Journal of the Medical Sciences for 
August, 1892, p. 167, Dr. Victor C. Vaughan details EXAMINATION OF DRINKING-WATER. 
219 
the method used in the Hygienic Laboratory of Michi- 
gan University. This method consists briefly of the 
following steps : — 
First. The water is collected in sterilized bottles. 
Second. Beef-tea tubes are inoculated with water, 
from one drop to one cubic centimeter being added to 
each tube. 
Third. The inoculated tubes are kept at 38° C. for 
twenty-four hours. 
Fourth. Many germs found in drinking-water will 
not grow at so high a temperature. Waters containing 
only such germs are pronounced safe. Certainly it 
must be admitted that a germ which will not grow at 
the temperature of the human body cannot possibly 
induce disease. 
Fifth. Of the germs which do grow at 38° or higher, 
some are toxicogenic to animals, while others are not. 
Whether the germs in a given sample are toxicogenic or 
not is determined by injecting the prepared cultures 
subcutaneously or intra-abdominally in rats, guinea-pigs, 
rabbits, or mice. 
Sixth. Waters containing only the non-toxicogenic 
germs growing at 38° or higher are reported as safe. 
Seventh. Waters containing toxicogenic germs are 
condemned. In testing the virulence, a germ is not 
pronounced toxicogenic unless it be found growing and 
multiplying in the animal inoculated with it. 
This method has been employed in the Hygienic 
Laboratory of the University of Michigan since Oc- 
tober, 1888. GLOSSARY. 
Taken and Abridged from Gould’s “ Illustrated Dictionary of 
Medicine, Biology and Allied Sciences.” 
A 
Aberration (ab-er-a'-shun). In optics, any imperfection of local- 
ization or refraction of a lens. A., Chromatic, A., Spherical. 
Acetonuria (as-et-on-u'-re-ah). The presence of acetone in the 
urine. 
Achromatic (ah-kro-mal'-ik). A. Lens, one without color, ex- 
actly neutralized by another lens having the same curvature but 
of unequal refractive index. 
Achromatin (ah-kro'-mat-in). The substance in the nucleus of a 
cell prior to division. So called because not readily stained by 
coloring agents. 
Actinospora chartarum (ak-tin-o-spo'-rah char-la'-runi). A 
parasitic fungus developing on paper and books. 
Adenogenesis (ad-en-o-jen'-es-is). The development of a gland. 
Adrenals (ad-re'-nalz). The supra-renal capsules. 
Adventitia (ad-ven-tish'-e-ah). The external covering or coat of 
the blood-vessels. 
Aerobic (a-er-o'-bik). Requiring oxygen (air) in order to live. A 
term applied to those bacteria requiring free oxygen. 
Agar-agar (a'-gar-a'-gar). A kind of glue made from certain sea- 
weeds, used in bacteriological studies to make a solution in 
which microorganisms are bred or kept. 
Albinuria (al-bin-u'-re-ah). I. Chyluria; whiteness of the urine. 
2. Albuminuria. 
Albuminuria (al-bu-min-u'-re-ah). The presence in the urine of 
albumin, a mixture of serum albumin and serum-globulin in 
various proportions. 
221 222 
MEDICAL MICROSCOPY. 
Allotoxin (al-o-toks'-in). Any substance, produced by tissue meta- 
morphosis within the organism, that tends to shield the body by 
destroying microbes or toxins that are inimical to it. 
Ameba or Amoeba (am-e'-bah). A colorless, single-celled, jelly- 
like protoplasmic organism constantly undergoing changes of 
form, and nourishing itself by surrounding objects. A. coli, 
ameba of dysentery. 
Amicrobic (ah-mi-kro'-bik). Not due to, or associated with, 
microbes. 
Amyloid (am'-il-oid). Starch-like pathological products. 
Anaerobic (an-a-e-ro'-bik). A term used of microorganisms that 
live in the absence of free oxygen or air. 
Antimycotic (an-te-mi-kot'-ik). Destructive of fungal micro- 
organisms. 
Antitoxic (an-te-toks'-ik'). Antidotal; counteracting poisons. 
Antitoxin (an te-toks'-in). A substance formed in the body of 
animals, either naturally or in consequence of inoculation with 
some pathogenic bacteria, that neutralizes the toxic products of 
these organisms. 
Aplanatic (ah-plan-at'-ik). Not wandering; rectilinear. A. 
Lens, a lens corrected for all aberration of light and color. 
A rectilinear lens. 
Apochromatic [ap-o-kro-mat'-ik). Without color. A. Lens, a 
lens for microscopic and optical purposes, with high correction 
of spherical and chromatic aberrations, and better “ definition.” 
A. Objective. 
Aseptic (ah-sep'-tik). Free from pathogenic bacteria, or septic 
matter. 
Autotoxic (aw-to-toks'-ik). Self-empoisonment through the ab- 
sorption of noxious products of katabolism, as in uremia. 
Azotemia (az-o-te'-ine-ah'). The presence of nitrogenous com- 
pounds in the blood; uremia. 
Azoturia (az-o-tu'-re-ah). An increase of the urea and urates in 
the urine. 
B 
Bacillar, or Bacillary (bas'-il-ar; bas'-il-a-re). I. Relating to 
bacilli or to a bacillus. 2. Consisting of or containing rods. 
Bacteriology (bak-te-re-ol'-o-je). That department of science that 
is concerned with the study of bacteria. GLOSSARY. 
223 
Bacterium [bak-te'-re-um). I. A term used to distinguish in a 
general way some of the simplest microscopic fungi, the Bac- 
teriacece, Fission-fungi, or Schizomycetes, and other closely 
allied microbes. 2. A genus of short, cylindrical, motile 
Fission-fungi. 
Binocular (bin-ok'-u-lar). In optics an instrument with two eye- 
pieces for use with both eyes at once. B. Vision, the faculty 
of using both eyes synchronously and without diplopia. 
Biogenesis [bi-o-jen'-es-is). The doctrine that living things are 
produced only from living things—the reverse of abiogenesis. 
Biologos [bi-olog'-os). A designation proposed for the intelligent 
living power displayed in cellular and organic action and reac- 
tion. 
Biology (bi-ol'-o-je). The science embracing the structure, func- 
tion, and organization of life-forms. 
Bioplasm (bi'-o-plazm). Any living matter, but especially ger- 
minal or forming matter; matter possessing reproductive vitality. 
Bioplast (bi'-o-plast). A mass or cell of bioplasm which is a unit 
of living matter. 
Blastema (blas-te'-mah). The formative lymph or pabulum of 
capillary exudation. A synonym of protoplasm. 
C 
Cacemia (kas-e'-me-ah, or kak-e'-me-afi). An ill-condition of the 
blood; depravity of the blood. 
Cachexia (kak eks'-e-ah). A term used to designate any morbid 
tendency, dyscrasia, depraved condition of general nutrition, or 
impoverishment of the blood, etc. 
Celloidin (sel-oid'-in). A concentrated form of collodion for use 
in imbedding objects for histologic purposes. 
Charlatan [shar'-lat-an). A quack; a pretender to medical skill; 
an advertising doctor. 
Chromatin {kro'-mat-in). The tingible portion of the protoplasm, 
forming a delicate reticular network of fibrils permeating the 
achromatin of a typical cell in process of division. It is 
called also Karyomiton. 
Chromation [kro-tna'-shun). The process of tingeing or staining. 
Chromatism [kro'-mat-izm). i. Abnormal coloration of any 
tissue. 2. Chromatic aberration. 224 
MEDICAL MICROSCOPY. 
Chromidrosis (kro-mid-ro'-sis). A rare condition of the sweat in 
which it is variously colored, being bluish, blackish, reddish, 
greenish, or yellowish. 
Chyluria (ki-lu'-re-afi). The passage of milky-colored urine. 
Cladothrix (klad'-o-thriks). A genus of bacteria, having long 
filaments, in pseudo-ramifications, with true spores. 
Coccidia (kok-sid'-e-afi). The so-called psorospermice—minute 
oval structures about 0.035 mm- long, with a thick capsule and 
coarsely granular contents. The organism is more properly 
called Coccidium oviforme, while the spores that it forms are 
termed psorospermise. 
Contagion (kon-ta' -jun). The process by which a specific disease 
is communicated between persons, either by direct contact or 
by means of an intermediate agent. Also the specific germ or 
virus from which a communicable disease develops. 
Contagium [kon-ta'-je-uni). Any virus or morbific matter by 
means of which a communicable disease is transmitted from 
the sick to the well; any living vegetable or animal organism 
that causes the spread of an infectious disease. 
D 
Desmobacterium [des-mo-bak-te'-re-um)\_pl. Desmobacteria~\. A 
group of microbes, so-called by Cohn, corresponding to the 
genus Bacillus of Klein. 
Diplococcus (dip-lo-kok'-us). A micrococcus whose spherules 
are joined two and two. 
E 
Eosinophile [e-o-sin'-o-fil). In bacteriology and histology, applied 
to microbes or histologic elements showing a peculiar affinity 
for eosin-stain. 
Epithelioma [ep-e-the-le-o'-mak). Carcinoma involving skin or 
mucous membrane. 
F 
Fuchsin (f00k'-sin) [after Leonhard Fuchs], Cj0H10N3. 
Fungicide (fun'-jis-ld). 1. Destructive to fungi; bactericide. 
2. An agent that destroys fungi or bacteria. GLOSSARY. 
225 
G 
Gemmation (jem-a' -shun). In biology, asexual reproduction by 
budding, as distinguished from fission and free-cell formation. 
Genesis (jen'-es-is). The act of begetting; development; origin; 
formation; generation. 
Genetic (jen-et'-ik). Pertaining to generation, or to anything 
inherited. G. Affinity, relationship by direct descent. 
Germ (jerm). In biology, (a) a portion of matter potentially vital 
and having within itself the tendency to assume a definite living 
form; a spore, a seed, an embryo. (<b) A microbe or bacterium. 
Germicide (jer'-tnis-id). A microbicide; an agent that destroys 
germs. 
Germiculture (jer'-me-kul-chur). The artificial culture of bacteria. 
Gonococcus (gon-o-kok'-us). A microbe, the specific cause of 
gonorrhea. 
H 
Hemameba (hem-am-e'-bak). A white blood-cell, so called from 
its resemblance to an ameba. 
Hematoblast (hem'-at-o-blast). Blood-plate; a rudimentary or 
immature red blood corpuscle. 
Hematocyst (hem'-at-o-sist). A cyst containing blood. 
Hematozoon (hem-at-o-zo'-on). Any living organism or animal 
in the blood. 
Hematuresis (hem-at-u-re'-sis). The passage of bloody urine. 
Hemocyte (hem'-o-slt). One of the protistan organisms found in 
the blood of man and animals, e. g., the parasite of malarial 
fever. 
Heteroblastic (het-er-o-blas'-tik). In biology, arising from a dif- 
ferent or abnormal source. 
Histopathology (kis-to-path-ol'-o-je). The study of minute 
pathologic changes or states. 
Hydruria (hi-dru'-re-ah). Excessive excretion of water by the 
kidneys. 
Hyperplasia (hi-per-pla'-ze-ah). The excessive deposit or aug- 
mentation of the elements of the tissue composing an organ. 226 
MEDICAL MICROSCOPY. 
I 
Immersion (im-trier'-shun). I., Homogeneous, a fluid between 
the objective of a microscope and the cover glass, having 
about the same refractive and dispersive power as the glass. 
I., Objective, a microscope-objective, usually of high power, 
the lower lens of which is immersed in a drop of water, gly- 
cerin, or oil, placed on the cover-glass of the object under ex- 
amination. 
Immune (im-mun/) [immunis, safe]. I. Safe from attack; pro- 
tected by vaccination, or some analogous procedure, or by 
previous illness. 2. A person who is protected against any 
special virus. 
K 
Karyokinetic [kar-e-o-kin-et'-ik). In biology, applied to the 
active stages of nuclei. 
Karyomitosis (kar-e-o-mit-o'-sis). The division or splitting of 
the nuclear mass of chromatin-fibers. 
L 
Latericious, Lateritious [lat-er-ish'-us). Pertaining to an uri- 
nary sediment resembling brick-dust. 
Leukoblast (lu'-ko-blast). I. The germ of a leukocyte; also, a 
leukocyte itself. 2. A cell in bone-marrow, of a type that is 
believed to become developed into a red blood corpuscle. 
Leukocyte (lu'-koslt). The colorless or white corpuscle of the 
blood. 
Leukomain, or Leucomain (lu-ko'-ttia-in). The nitrogenous 
bases or alkaloids necessarily and normally developed by the 
vital functions or metabolic activity of living organisms, as dis- 
tinguished from the alkaloids developed in dead bodies, and 
called ptomains. 
Lipogenous (lip-oj'-en-us). Fat producing. 
Lipuria (lip-u'-re-ah). The presence of fat in the urine. 
Lithuria {lith-u'-re-ah). A condition marked by excess of lithic 
acid, or its salts, in the urine. 
Lymphocyte (lim'-fo-slt). 1. A lymph-cell. 2. One of Ehrlich’s 
classes of leukocytes, comprising those small cells having large 
nuclei and a very small amount of protoplasm. GLOSSARY. 
227 
M 
Macrochemistry (mak-ro-kem'-is-tre). Chemistry in which the 
reactions are observable with the naked eye. 
Macroscopic {tnak-ro-skop'-ik). Large enough to be seen by the 
naked eye; gross; not microscopic. M., Morbid Anatomy, 
naked-eye or gross morbid anatomy. 
Megalocyte {nieg' al-o-sit). A red blood-corpuscle larger than 
the average; especially the form that characterizes pernicious 
anemia. 
Mesobacteria (mez-o-bak-te'-re-ah). Medium-sized vegetable mi- 
croorganisms. 
Microbe (mi'- krob). The generic name for microorganisms, 
whether animal or vegetable. In ordinary use the term microbe 
is equivalent to schizomycete, and designates a vegetable micro- 
organism. 
Microbicide (mi-kro'-bis-td). I. Destructive to microbes. 2. An 
agent that destroys microbes. 
Microbiology hni-kro-bi-ol'-o-je). Bacteriology, or the science of 
microorganisms. 
Micro-chemistry (tni-kro-kemf-is-tre). The chemic investigation 
of the more minute substances of nature. 
Micrococcus (mi-kro-kok'-us). A genus of schizomycetous micro- 
organisms, having spheric elements, isolated, united in two’s or 
in larger numbers, or disposed in chaplets, or masses of zooglea. 
Sometimes they are united in such a way as to resemble a bunch 
of grapes and are then called Staphylococci. When united 
in couples they are called Diplococci. If arranged in strings 
or chaplets they are called Streptococci. 
Micron (vii'-kron). The millionth part of a meter or a thousandth 
of a millimeter. It is the equivalent of of an English 
inch, and its symbol is fi. 
Microphyte [mir-kro-fit). Any microscopic plant, especially one 
that is parasitic in habits. 
Microscopist (rui-kros'-ko-pist). One who is skilled or expert in 
the use of the microscope. 
Microscopy (mi-krosf-ko-pe). The use of the microscope; micro- 
scopic study or observation. 
Microtomist (nii-krot'-o-mist). One who cuts sections with the 
microtome. 228 
MEDICAL MICROSCOPY. 
Monochromatic (mon-o-kro-mat'-ik). Having but one color. 
Mordant (mor'-dan/) \jnordere, to bite]. A substance that fixes 
the dyes used in coloring textures, or in staining tissues and 
bacteria. 
Mucinoid (mu'-sin-oid). Resembling mucin. 
Mucinuria (mu-sin-uf-re-ah). The presence of mucin in the urine. 
Mucous (tnu'-kus). A term applied to those tissues that secrete 
mucus. 
Mucus (mu'-kus) [L.]. The viscid liquid secretion of mucous 
membranes, composed essentially of mucin holding in suspen- 
sion desquamated epithelial cells, leukocytes, etc. 
N 
Neoplasm, Neoplasma (ne'-o-plazm, ne-o-plaz'-mah). A new 
growth of tissue marked by histologic difference from its matrix; 
a tumor. 
Neuroplasm (nu'-ro-plazm). That form of bioplasm exhibited in 
living brain-tissue. 
Nidus (ni'-dus) [L., nest], A central point or focus of infection ; 
a place in which an organism finds conditions suitable for 
growth and development. 
Nigrosin (nig'-ro-sin). A blue-black anilin-dye, useful in stain- 
ing sections of brain-tissue. 
Nosogeny (nos-oj'-en-e). The development and progress of 
diseases. 
Nosomycosis (nos-o-mi-ko'-sis). Any disease due to the presence 
of a parasitic fungus, or schizomycete. 
Nucleolus (nu-kle'-o-lus). The small spheric body within the 
cell-nucleus. 
Nucleus (nu'-kle-us). The essential part of a typical cell, usually 
round in outline, and situated near the center. 
O 
Oidium (o-id'-e-um) [dim. of uov, egg]. A genus of parasitic 
fungi. O. albicans is found in thrush, upon the tongue. O. 
lactis, the white mold found on milk, bread, etc. 
Organism (or'-gan-izm). A living being, animal or vegetable, 
simple or composed of many organs; also the assemblage of GLOSSARY. 
229 
organs constituting a living being. O., Micro-, a minute or 
microscopic body or organism; a schizomycete; a bacterium. 
Organoid (or'-gan-oid). A term applied to tumors composed of 
several tissues and resembling an organ, as carcinoma, which 
somewhat resembles an epithelial gland. 
Orgasm [or'-gasm). The crisis of venereal passion. 
Osier’s Method. A method of studying blood-plaques; a drop 
of osmic acid is placed on the cleansed finger, which is then 
pricked and the drop transferred to a slide. 
Oxaluria [oks-al-u'-re-ali). A term used to indicate the presence 
of calcium oxalate in the urine in an undue amount. There is 
a white deposit on standing. It occurs in the urine of hypo- 
chondriacal and depressed patients, and in that of gouty 
patients. Excessive venery and masturbation will produce it, 
as also will the ingestion of certain foods, as rhubarb. 
P 
Paludism [pal'-u-dizm) [pains, a marsh]. Malarial poisoning; 
impaludism. 
Paraffin [par'-af-in). A white, odorless, translucent, crystalline 
hydrocarbon obtained from coal tar, or by the destructive dis- 
tillation of wood. In a pure state it resembles white wax in 
physical properties. 
Parasite [par'-as-it). In biology, an organism that inhabits 
another organism and obtains nourishment from it; it may be a 
phytoparasite or a zooparasite, an ectoparasite ox an endoparasite, 
occasional or constant, temporary or stationary, obligate or 
facultative, a true parasite or a pseudo-parasite. 
Parasiticide [par-as-it'-is-ld). Any substance destructive of 
parasites. 
Pathogenic, Pathogenetic [path-o-jen'-ik, path-o-jen-et'-ih). 
Producing disease. P. Microorganism, any one of the 
various forms of microbic life which, when introduced into 
the system, causes disease. 
Phagocyte [fag'-o-sit). Phagocytes are fixed—endothelial cells, 
fixed connective-tissue cells, and free—the wandering cells or 
leukocytes. In man the colorless blood-cells, as well as other 
kinds of cells, are credited with playing the role of phagocytes. 
They are believed to englobe wrecks of larval organs, degrada- 
tion-products or excretion products, foreign particles, schizo- 230 
MEDICAL MICROSCOPY. 
mycetes, hematozoa, etc.—their activity varying as the loga- 
rithm of the excitation. They digest the soluble parts and reject 
the insoluble residue. They play an active part in the meta- 
morphosis of tissues and organs, in inflammation, and as pro- 
phylactic agents. 
Phagosite (fag'-o-sit). An animal organism that feeds on but 
does not dwell within or on its host, e. g., the leech, lamprey, 
camel-tick, vampire bat. 
Phosphaturia (fos-fat u'-re-ah). A condition in which an excess 
of phosphates is passed in the urine. It can be diagnosticated 
by a quantitative analysis of the urine for phosphates by the 
uranium method. 
Photomicrograph [fo-to-mi'-kro-graf). A photograph of a small 
or microscopic object, usually made with the aid of a micro- 
scope, and of sufficient size for observation wdth the naked eye. 
Photuria (fo-tu'-re-ah). Phosphorescence of the urine. 
Poikilocyte (poi'-kil-o sit). A large, irregularly shaped red 
blood-corpuscle. Poikilocytes are most abundant in the blood 
in pernicious anemia, but also occur in other forms of anemia. 
Polyuria {pol-e-u'-re-ah). Excessive secretion of urine. The 
causes of temporary polyuria are, excessive ingestion of fluids, 
cold, suppression of perspiration, the use of diuretics; it occurs 
in the crisis of fevers, and in certain neurotic conditions, as 
hysteria, and in nervous excitement. A permanent polyuria is 
met with in diabetes mellitus, diabetes insipidus, chronic inter- 
stitial nephritis, and in amyloid disease of the kidneys. 
Protozoa (pro-to-zo'-ah). The lowest class of the animal king- 
dom, comprising organisms which consist of simple cells or 
colonies of cells, and which possess no nervous system, and no 
circulatory organs. 
Ptomain (to'-ma-in). Any one of the active, inanimate septic or 
toxic substances resulting from processes of decomposition and 
disintegration of albuminous materials. Ptomains have been 
found in foods, as in mussels, oysters, eels, sausage, ham, 
canned meats, cheese, milk, ice-cream, etc. The pathogenic 
action of certain bacteria may be due to their production of 
ptomains. 
Pyemia (pi-e'-me-ah). Phlebitic septicemia, with the presence of 
pyogenic microorganisms in the blood and with the formation 
wherever they lodge of secondary embolic or metastatic ab- 
scesses. GLOSSARY. 
231 
Pyogenic (pi-o-jen'-ik). Producing or relating to pus-formation. 
Pyrotoxin (pi-ro-toks'-in). A toxic agent generated in the course 
of the febrile process. 
Pyuria (pi-u'-re-ah). Pus in the urine. 
Q 
Quack (kwak). One who practises quackery; a pretender to 
medical skill; a peddler of his own medicines and salves. 
R 
Reagent (re-a'-jent). In chemistry, anything used to produce a 
reaction, or to test for the presence of an element. A test. 
Rhabdomyoma (rab-do-mi-o'-mah). A rare form of myoma 
characterized by the presence of striated muscular fiber. 
S 
Sanatorium [san-at-o'-re-uni) [sanare, to heal]. An establish- 
ment for the treatment of the sick ; especially a private hospital. 
Sanitarium (san-it-a'-re-um) \sanitas, health]. A health station. 
A place or institution where the conditions are such as espe- 
cially to promote health and vigor. The word is often incor- 
rectly employed for sanatorium, which is a hospital or place for 
curing those who are sick. A sanitarium may be used as a 
sanatorium but it is not necessarily the same thing. 
Saprogenic (sap-ro-jen'-ik). Causing putrefaction; caused by 
putrefaction. 
Saprophytic (sap-ro-ft'-ik). In biology, growing on or in decay- 
ing organic matter. 
Sarcina (sar-si'-nah). A genus of Schizomycetes, or bacteria, 
having spheric or ovoid cells dividing in three directions, thus 
producing cubic masses of greater or less size. 
Schizomycetes (skiz-o-mi-se'-tez). In biology, an order of Fungi; 
the so-called Fission-fungi or Bacteria. 
Schizomycosis (skiz-o-mi-ko'-sis). A disease due to schizomy- 
cetes. 
Scirrhus, or Scirrus (skir'-us or sir'-us). A scirrhous or hard 
carcinoma. 232 
MEDICAL MICROSCOPY. 
Septic (sep'-tik). Relating to putrefaction. S. Infection, infec- 
tion with pathogenic microorganisms. S. Intoxication, ab- 
sorption of septic matter. S. Pestilence. 
Septicemia, Septicaemia {sep-tis-e'-me-ah). A condition induced 
by the absorption of septic products. Pyemia is septicemia plus 
the formation of secondary or embolic abscesses. 
Septico-pyemia (sep-tik-o-pi-e'-me-ah). The condition of com- 
bined septicemia and pyemia; septic and purulent infection. 
Septodiarrhea (sep-lo-di-ar e'-ak). Septic diarrhea. 
Septodysenteria (sep-to-dis-en-ter'-e-ah). Septic dysentery. 
Sequela (se-kivel'-ah). The consequence or abnormal condition 
following an injury or the abatement of a disease ; any diseased 
or abnormal condition that follows an attack of disease or an 
injury. 
Seralbumin (slr-alf-bu-min). Serum-albumin ; the albumin found 
in the blood, in distinction from that of the egg, ovalbumin. 
Sewage (su'-aj). The heterogeneous substances constituting the 
excreta and waste matter of domestic economy and the con- 
tents of drains. 
Sewerage (su'-er-aj). The collection and removal of sewage. 
Society Screw. The screw at the lower end of the draw-tube or 
body-tube of a microscope for receiving the objective. 
Streptobacteria (strep-to-bak-te'-re-aK). In biology, short, rod- 
shaped bacteria associated in chains. 
Streptococcus (strep-to-kok'-kus). A genus of coccaceous schizo- 
mycetes, of which the cocci are arranged in strings or chaplets. 
Many of the species are believed to be pathogenic. 
Streptothrix (strep-loth'-riks). In biology, a genus of Schizomycetes, 
the cells uniting into simple or branching threads. 
Stroma (stro'-mah). The tissue forming the substratum or frame- 
work upon which the essential structures of an organ rest. 
Substage (sub'-staj). The arrangement beneath the stage of a 
microscope for the diaphragms, condenser, illuminator, and 
other accessories. 
Supernatant (su-per-na'-tant). Floating upon the surface of a 
liquid. 
T 
Tabescence (tab-es'-ens) \tabes, wasting]. Wasting; marasmus; 
emaciation. GLOSSARY. 
233 
Telangioma (tel-an-je-o'-malt). A tumor composed of dilated 
capillaries. 
Teniacide, Taeniacide (te'-ne-as-id). Destructive of tapeworms; 
a remedy that destroys tapeworms. 
Teniafuge, Taeniafuge (te'-ne-af-uj). An agent that expels, 
without necessarily killing, tenia. 
Teratism (ter'-at-izm). Any anomaly of conformation, whether 
congenital or acquired through disease or injury. 
Teratoma (ter-at-o'-mah). A congenital tumor, which may con- 
tain various concretions of organic tissue, as teeth, hair, and 
other erratic material. Dermoid cyst. 
Toxemia (toks-e'-me-ah). A condition of the blood in which it 
contains poisonous products, either those produced by the body- 
cells and not properly eliminated, or those due to the growth of 
microorganisms. 
Toxicogenic (toks-ik-o-jen'-ik). Giving rise to poisons; produc- 
ing a toxic substance, as a toxicogenic microorganism. 
Trabecula (tra-bek'-u-lah). Any fibrous process, layer, or cord 
which goes to make up a framework in an organ or viscus. 
Traumatism (traiv'-mat-izm). The condition of one suffering 
from injury. The systemic condition following trauma. 
Trichina (trik-i'-nah, or trik-e'-na). A genus of parasitic nema- 
tode worms. 
Trichinosis (trik-in-o'-sis). A disease produced by the ingestion 
of meat, pork, or sausage, containing the Trichina spiralis. 
Typhotoxin (li-fo-toks' in). A ptomain discovered by Brieger, 
and believed to be the special product of the Koch-Eberth 
typhoid-bacillus. 
Tyrotoxicon (ti-ro-toks'-ik-on). A ptomain obtained by Vaughan 
from poisonous cheese, poisonous milk, poisonous ice-cream, 
etc. 
U 
Urinalysis (u-rin-al' -is-is). The analysis of the urine. 
Urinology (u-rin-ol'-o je). The science of the analysis and 
diagnostic significance of urine. 
Uroscopist (u-ros'-ko-pist). One who makes a specialty of urin- 
ary examinations. 234 
MEDICAL MICROSCOPY. 
V 
Verrucose, Verrucous (ver'-u koz, ver' u kus). Warty; covered 
with or having warts. 
Vestigium (ves tij'-e um). An anatomic relic of fetal or embryonic 
life. Thus, the thymus gland becomes in adults a vestigium 
Virus (vi'-rus) [L.]. A poison that causes a morbid process or 
disease; any pathogenic microbe. 
Viscous (vis'-kus). Glutinous, ropy, sticky. 
Viscus (vis' kus) [L.: pi., Viscera]. Any organ enclosed within 
either of the four great cavities, the cranium, thorax, abdominal 
cavity, or pelvis; as the brain, intestine, spleen, bladder, uterus, 
lungs, liver, etc. 
W 
Wilks’s Kidney. The large white kidney of chronic parenchy- 
matous nephritis. 
X 
Xylol (zi'-lol). Dimethyl benzene. A volatile hydrocarbon some- 
what resembling benzol. 
Z 
Zooglea (zo o gle'-ah). In biology, a stage in the life-history of 
certain Schizomycetes, or bacteria, in which they lie embedded 
in a gelatinous matrix secreted by the microbes themselves. 
Zymotic (zi tnot'-ik). In biology, pertaining to zymosis, or fer- 
mentative changes produced by an organized ferment, or zyme. INDEX. 
ABERRATION, chromatic, 45 
spherical, 45 
Achromatic condenser, 52 
Actinomycosis, 139 
Adenoma, 147 
Amceba coli, 130 
Angeioma, 150 
Angle of aperture, 46 
Animal parasites, 210 
Asiatic cholera, 95 
Bacillus anthracis, 129 
of asiatic cholera, 101 
of cholera nostras, 102 
of diphtheria, 120 
of glanders, 125 
of influenza,128 
of leprosy, 126 
of scarlet fever, 121 
of syphilis, 123 
of tetanus, 127 
of tubercle, 112 
of typhoid fever, 104 
Bacteriology, 89 
Benzol, uses of, 108 
Blood, the, 201 
corpuscles in urine, 173 
counting of, 204 
white, 206 
platelets, 202 
Books, list of, 54, 55, 56 
Borax-carmin, 77 
methyl-blue, 83 
Bright’s disease, 181 
Carbolized double stain, 85 
Carcinoma, 155 
parasitic theory of, 161 
Celloidin embedding, 67 
Centering diagram, 75 
Charcot-Leyden crystals, 212 
Cholesterin, 200 
Chondroma, 145 
Chylous urine, 196 
Clean slides and covers, 60 
Colloid cancer, 160 
Comma bacillus, 101 
Cotton fibers, 57 
“ Continental ” stands, 39, 41 
Cover-glass preparations, 100 
Croupous pneumonia, 117 
Cultivation-media, 99 
Cutting sections, 70 
Cylindroids, 217 
Cystin, 197 
Damp chamber, too 
Decalcifying mixture, 64 
Defining power, 45 
Diaphragms, 52 
Double nose-piece, 53 
Double staining, 78 
Egg albumin, solution of, 74 
Embedding in paraffin, 66 
Encephaloid cancer, 157 
Epithelioma, 159 
cylinder-celled, 158 
flat-celled, 159 
Extraneous matters, 58 
Exudates, 211 
Eye-piece, 42 
micrometer, 50 
False casts, 177 
Fecal discharges, 213 
Fibromata, 143 
Finkler-Prior bacillus, 102 
Fixing and hardening, 68 
sections on the slide, 74 
235 236 
INDEX. 
Flatness of field, 45 
Fleming’s method, 87 
Forms of bacteria, 92 
Formulary, 81 
Freezing microtome, 73 
Fungi in urine, 183 
Gibbes’ method, 85 
Giant-celled sarcoma, 153 
Glioma, 155 
Gonococcus, 122 
Gram’s method, 87 
modification of, 87 
solution, 87 
Granular casts, 187 
Gray, Dr. Wm. M., 76 
Hardening tissues, 63 
Hematoxylin, 79 
double stain, 80 
Hemin crystals, 207 
Hemocytometer, 204 
Hippuric acid, 198 
How to work, 54 
Hyaline casts, 187 
Illumination, 51 
Immersion objectives, 47 
Indigo-carmin, 79 
Interstitial embedding, 66 
Intestinal and other parasites, 213-16 
Karyokinesis, 87 
Killing and hardening, 63 
Leiomyoma, 148 
Lens, oil-immersion, 48 
Leucin, 199 
Leukocytes, 175 
Leukocythemia, 203 
Leukocytosis, 203 
Leukemia, 208 
Linear magnifying powers, 49 
Lipomata, 145 
Loffler’s solution, 83 
Lymphangeiomata, 150 
Lymphomata, 148 
Making casts, 69 
Melanotic cancer, 160 
sarcoma, 154 
Micrococcus urese, 183 
Micrometer, eye-piece, 50 
Microorganisms, 95 
in food and drinks, 218 
in the skin, 217 
Microscope, choice of, 40 
use of, 40 
Microtomes, 71, 72 
Mirror, 51 
Mixed cell sarcoma, 154 
Molds for paraffin casts, 68 
Mucous cancer, 160 
Myomata, 148 
Necessary outfit, 34 
Neoplasms, 142 
Neuromata, 149 
Nutrient agar-agar, 99 
gelatin, 97 
Objectives, 44 
Organized urinary deposits, 177 
Osteomata, 147 
Oxalate lime crystals, 192 
Papillomata, 147 
Paraffin, 65 
Penetrating power, 46 
Plasmodium malariae, 133 
Plate-cultures, 98 
Pneumonococcus, 119 
Poikilocytosis, 208 
Potato culture-medium, 106 
Psammoma, 155 
Pulmonary tuberculosis, 109 
Pus, Donne’s test, 176 
Ray fungus, 139 
Relapsing fever, 138 
Renal epithelium, 170 
Renal tube-casts, 187 
Resolving power, 46 
Round-celled sarcoma, 152 
Salt solution, 64 
Sarcomata, 151 
Schizomycetes, 93 
Scirrhus cancer, 157 
Section cutting, 70 
flattener, 74 
Seminal fluid, 188 
Sharpening microtome knife, 74 
Spermatozoids, 188 
“ Spider-cells,” 104 Spindle-celled sarcoma, 152 
Sputum, in, 210 
Staining sections, 77 
Sub-stage accessories, 52 
Taenia saginata, 215 
solium, 214 
Teratoma, 155 
Trichina spiralis, 216 
Triple phosphates, 193 
Tubercle bacilli, m 
in urine, 189 
Tuberculous infection, 114 
Tumors, 143 
Tyrosin, 199 
Urates, 189 
Urinary examinations, 163 
INDEX. 
237 
Urine, composition of, 165 
red blood-corpuscles in, 173 
Uric acid crystals, 191 
Vibrio proteus, 103 
Vaginal epithelium, 170 
Vesical epithelium, 170 
Waste of material, 62 
Water bath and oven, 66 
Water examination, 218 
Whipworm, 216 
Woolen fibers, 57 
Working-distance, 47 
Xanthin, 198 
Xylol, 59, 117 THE WORKS OF JAMES TYSON, M. D, 
Professor of Clinical Medicine in the University of Pennsylvania 
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