Extracted from the American Journal of the Medical Sciences for January, 1881. 
ON THE ACTION OF 
CARBOLIC ACID UPON CILIATED CELLS AND 
WHITE BLOOD CELLS. 
BY 
T. MITCHELL PRUDDEN, M. D., 
Lecturer on Normal Histology in the Medical Department of Yale College, Assistant in the Pathological 
Laboratory of the Alumni Association of the College of Physicians and Surgeons, New York. 
COMPLIMENTARY. [Extracted from the American Journal of the Medical Sciences for January, 1881.] 
ON THE ACTION OF CARBOLIC ACID UPON CILIATED CELLS 
AND WHITE BLOOD CELLS. 
Br T. MITCHELL PRUDDEN, M.D., 
LECTURER ON NORMAL HISTOLOGY IN THE MEDICAL DEPARTMENT OF YALE COLLEGE, ASSISTANT 
IN THE PATHOLOGICAL LABORATORY OF THE ALUMNI ASSOCIATION OF THE 
COLLEGE OF PHYSICIANS AND SURGEONS, NEW YORK. 
Rich as is the literature of the last two decades in records of investiga- 
tions on the local and general action of carbolic acid upon the living or- 
ganism, the minute changes which it produces in the cells themselves, 
seems to have attracted but little attention. To those who have adopted 
the postulate upon which the Listerian treatment of wounds is based, the 
study of the action of the antiseptic upon bacteria and allied organisms 
has seemed very naturally of paramount importance ; while those, on the 
other hand, who denying, or at least not accepting, as yet proven, the the- 
oretical considerations in which the Listerian treatment centres, but yet 
recognizing fully the value of carbolic acid as a topical application, seem 
to have contented themselves for the most part with very general explana- 
tions of its favourable action. 
The significant fact that, other things being equal (sanitary conditions, 
care, etc.), the strict adherence to the Listerian doctrine and details, has, 
with perhaps the exception of certain classes of operations (ovariotomy, 
etc.), given results which, so far as statistical data are reliable, seem to be 
in no wise superior to those obtained by surgeons who use carbolic acid as 
a dressing agent, but without inflexible adherence to the details insisted 
upon by Lister, and which, if his postulate be true, would appear indispen- 
sable, has suggested the query, whether, after all, the beneficial effect of 
carbolic acid, which in some measure all acknowledge, may not really be 
largely due to its direct action upon the tissues of the healing wound. This 
view, which so far as it was formulated seems to have been the current 
one before the action of micro-organisms in wounds was postulated and 
became popular, was persistently urged by Beale1 in 1872 on theoretical 
grounds, and has been ably advocated from the standpoint of the practical 
surgeon by Prof. T. M. Markoe,1 in a recent number of this Journal. 
* Beale, Disease Germs, Their Nature and Origin, 2d ed., 1872, p. 279, et seq. 
3 Markoe, Through Drainage in the Treatment of Open Wounds, Am. Journ. Med. 
Sci., April, 1880. 
1 2 
In the hope that some light might be thrown upon this important ques- 
tion by observations upon the action of carbolic acid on some of the ele- 
mentary organisms of living tissues, a series of experiments was instituted 
the results of which it is the purpose of the present paper to record. The 
question proposed for solution was : What effects, if any, are produced by 
solutions of carbolic acid of varying strength, first upon the structure, and 
second upon the function of certain living cells? The two varieties of cells 
selected for this purpose were ciliated cells and white blood cells, and these 
were chosen because with a tolerably well-known structure one of the func- 
tional expressions of their life is in well-defined and easily studied move- 
ments. 
The preparation used was Merck’s purified carbolic acid, in crystals, 
and this was in all cases dissolved in a half per cent, solution of sodium 
chloride ; the latter solution acting in most cases towards living tissues as 
an approximately indifferent fluid, or its effects being at least well known 
or determinate. Solutions were used of the strength of 1-20, 1-50, 
1-100, 1-400, 1-800, 1-3200. The most extended series of ex- 
periments were made upon both ciliated and white blood cells from the 
frog (Rana esculenta), while less extended experiments were made upon 
both kinds of cells from the rabbit and upon the white blood cells of man. 
It does not seem desirable to record at length the individual experiments, 
but simply to describe the methods employed, and give their results. In 
each case a sufficient number of experiments vvas made to insure the elim- 
ination of such errors as isolated or a too limited number of observations 
in studies on the living tissues are always liable to lead, owing to varying 
conditions of vitality over which the investigator has no control, and of 
whose nature or presence he often may not even be aware. Control ex- 
periments, in which the indifferent fluid alone was used, were also made 
in all cases in which the character of the observation made them desirable. 
The Action of Carbolic Acid upon Ciliated Cells of the Frog Its effect 
was first studied upon living cells scraped from the palate, and second upon 
the cells in situ on the edge of the tongue of the living curarized animal, 
with the capillary circulation intact at their base. 
In the first series of experiments, the cells having been gently scraped 
from the mucous membrane, were teased apart on a slide in half per cent, 
salt solution, protected from pressure by two bits of thin blotting paper on 
either side, and covered as usual by thin glass. After bringing into the 
field suitable masses of cells, in which the ciliary movement was vigorous, 
the carbolic acid in different degrees of dilution was allowed to flow under 
the cover-glass and was drawn off on the other side by blotting paper in 
the usual way. In order to avoid such errors as might arise from the con- 
centration of the salt solution by evaporation, it is customary to inclose 
specimens exposed in this way to the action of fluids, with a rim of oil or 
paraffine ; but when the observation is to be long continued, such an ex- 3 
elusion of oxygen should be guarded against, and hence in these experi- 
ments a slow current of the irrigating fluids was kept constantly passing 
through beneath the cover-glass. It has been shown by Steinbuch, Pur- 
kinje and Valentin, Roth,1 and others, that mechanical influences, such as 
currents of fluid, are capable under certain conditions of accelerating cil- 
iary movement or even exciting it when it has once ceased. But a slow 
constant current being maintained in these experiments, alike with the 
indifferent and the carbolated fluid, this factor was probably made uniform 
for both. 
Under these conditions the effects produced by carbolic acid are briefly 
as follows: In the strength of 1—20, the ciliary movement ceases on the 
instant of contact with the cell and the cilia loose their well-defined con- 
tours, shrinking to a tangled mass of curled and twisted filaments, which 
soon gives place to a narrow band of irregular fine granules. The cell 
body becomes at once deeply cloudy, and then coarsely granular, shrink- 
ing rapidly, and the protoplasm is soon resolved into two distinct kinds of 
material; the one clear, transparent, and feebly refractive: the other, 
formed in varying quantity, strongly refractive and forming irregular 
coarse shining reticular masses within the cell, or appearing in the form of 
larger and smaller shining globules or droplets, which may or may not 
entirely conceal the nucleus. Specimens kept under observation for 35 
days showed within the first 48 hours some additional shrinkage, and a 
tendency to become brownish and dry in appearance, while the shining 
droplets usually became more or less coalescent and often freed themselves 
completely from the rest of the cell. After this time no marked change 
occurred in the cell within the above period. The nature of the shining 
globules and reticular masses developed from the protoplasm by carbolic 
acid, which are but partially soluble in alcohol and ether and not at all 
changed in colour by osmic acid or iodine, the writer was unable to 
discover. 
The action of solutions of 1-50 and 1-100 are closely similar. In a 
small proportion of cases, especially when the ciliary movement is at all 
languid, it becomes distinctly accelerated as the acid reaches the cells, and 
then, which is true for most of the cells at first* the movement becomes 
slower and usually ceases altogether in a few seconds, although in some 
cases, especially with the weaker solution, it may persist for several min- 
utes. As the vibration ceases, the cilia may be indistinct in outline, or 
curved, or twisted, or may stand out from the cell straight and stiff. Then 
occur degenerative changes in the protoplasm similar to those above de- 
scribed, though less rapidly developed. Shining globules appear in the 
protoplasm, the nucleus shrinks, not unfrequently exhibiting as it does so 
1 Steinbuch, Analecten neuer Beobachiungen und Untersuchungen fur die Xatur- 
kunde, 1802, p. 66. Purkinje and Valentin, De phenomeno motus vibratorii, etc. 
Wrastl., 1835. Roth, Virchow’s Archiv, Bd. 37, p. 191. 4 
distinct movements in the intra-nuclear network; and especially note- 
worthy in solutions of this strength, is the development in a large number 
of the cells, within a few minutes, of a wedge-shaped more or less strongly 
refractive, sharply outlined network, extending from the ciliary border 
to the nucleus, within which are sometimes seen delicate parallel lines, 
perpendicular to the free border of the cell. This peculiar network, lying 
invariably between the ciliary border and the nucleus and neverextending 
below the latter, would suggest a difference in the structure of the proto- 
plasm in this part of the cell which is not without significance in view of 
the present uncertainty of our knowledge concerning the relations between 
the cilia and the nucleus and cell body. Aside from a tendency of the 
shining globules to coalesce and gather outside the cell body within the 
first 48 hours, no marked change was noticed in cells exposed to the acid 
for 35 days. 
Under the action of solutions of 1-200, the vibration is, at first, in many 
cases, distinctly accelerated and then becomes slower, the cell body be- 
coming slightly cloudy, and ceases altogether as a rule within a few min- 
utes. But it must be said that under these as well as other conditions, 
certain cells or groups of cells resist the action of the acid much longer 
than others which are exposed, so far as can be determined, to precisely 
the same conditions. After the movement ceases the wedge-shaped sub- 
ciliary network as well as a few shining globules are sometimes slowly 
developed. But neither are as well marked or abundant as when solutions 
of greater concentration are used. Not infrequently, sometimes before the 
ciliary movement ceases, larger and smaller hyaline droplets may be seen 
exuding from the free border of the cell, often in great numbers.1 If in 
the use of solutions of the above strength, at the instant when the vibra- 
tions cease, the carbolated fluid is washed out by the salt solution they 
may, under favourable circumstances, recommence and continue for a few 
minutes but cease permanently, as a rule, in a short time. 
With solutions of from 1-400 to 1—800, the vibration is gradually 
brought to rest usually in from 60 to 90 m., the cilia standing out stiffly 
from the cell. The structural changes produced in the cell body are 
inconsiderable; they undergo the same sort of degeneration, but a little 
more slowly, observed in cells lying in simple salt solution. Here, too, if 
very soon after the cessation of movement the acid be washed out by salt 
solution, a restitution of the ciliary vibration often occurs, frequently con- 
tinuing for a considerable time. It may even be again suppressed by the 
acid, and once more re-established by the indifferent fluid. In still more 
dilute solutions, 1-1600 to 1-3200, no characteristic structural changes 
1 This appearance, the occurrence of which under other conditions Virchow described 
many years ago (Virchow’s Archiv, Bd. vi. p. 133), is instructive as showing how pro- 
found are the changes which cells may undergo without a suspension of functional 
activity. 5 
were observed, but while exposed to the action of the acid, the movement 
was as a rule somewhat slower and came sooner to rest than when salt 
solution alone was employed. 
A less extended series of observations upon the living ciliated cells from 
the air-passages of the rabbit showed that upon these the carbolic acid 
has essentially the same effect, but that they are somewhat more suscep- 
tible than those of the frog, both to influences causing structural changes 
and interference with functional movement. 
In the second series of experiments on the ciliated epithelium, in which 
the action of the acid on the cells in situ in the living animal was studied, 
the very convenient frog plate for the study of the tongue, devised by Prof. 
Richard Thoma, of Heidelberg, was employed.1 If upon this apparatus 
the tongue be stretched as tightly as is compatible with the maintenance of 
a good circulation, and then covered and suitably irrigated, a most admira- 
ble view may be obtained at its edges of the ciliary movement under nearly 
normal conditions. Although the structural details of the cell bodies in 
this picture are not readily made out, the movements of the cilia are very 
distinct, and conclusions drawn from their study rest upon a much firmer 
basis than those derived from a study of the teased cells alone, since the 
latter, in the preparation for the experiment, are placed under excessively 
abnormal conditions. Still each method has its distinct advantages, and 
neither can be altogether dispensed with. 
1 This exceedingly useful apparatus consists of a rectangular plate of brass 12 cm. 
long by 7 cm. wide, covered above by a thin sheet of hard rubber. The plate is 
pierced at one end by an opening 2 cm. by 1 cm., into which a polished strip of glass 
is cemented which is thicker at the outer than at the inner end, corresponding to the 
difference in thickness of the frog’s tongue at the tip and base. Around this glass 
plate over which the tongue of the curarized frog is gently stretched by pins driven 
into strips of cork on either side, a narrow rim of brass is fastened, making a gutter 
around the tongue from which two caoutchouc tubes lead off into a receptacle for the 
waste irrigating fluid under the table. On one side of the plate are two canula hold- 
ers, so arranged as to turn in all directions, by means of which the irrigating fluids 
may be carried through fine glass tubes to any part of the tongue or made to irrigate 
the whole. The irrigating fluid is supplied from a Mariotte bottle set at a suitable 
height above the level of the microscope stage upon which the frog plate is clamped 
in the usual way, and the whole so inclined that the waste irrigating fluid will readily 
flow off. 
The advantages of this apparatus are : that any surface exudation is washed away 
and the picture kept clean and clear; that the tongue may be kept for a very long 
time under observation, its proper moistening being automatical, and the waste fluids 
conveyed away without any danger of coming in contact with the microscope; that 
facilities are placed at the command of the investigator for varying in a multitude of 
ways the conditions under which the tissues are placed, the changes in irrigating 
fluids being effected simply by closing one clip and opening another. The tongue fur- 
thermore, notwithstanding its greater thickness at the base, is, owing to the obliquity 
of the glass plate upon which it rests, horizontally placed, and may be covered by thin 
glass, under which the fluids from the irrigating canulae can be directed and studied 
with high powers. This plate is fully described and figured by Thoma in Virchow’s 
Archiv, Bd. 65, p. 36. 6 
This series of experiments, between 40 and 50 in number, showed that 
in general the action of carbolic acid upon the ciliated cells in situ is 
essentially the same as upon the separated cells, except that the former 
are considerably more resistant to its action. The same degenerative 
changes in the protoplasm are produced with the stronger solutions, and 
with these as well as with those considerably dilute a tendency to desqua- 
mation is usually observed. The most noteworthy effect produced upon 
the cells under these conditions by dilute solutions, is their power of hold- 
ing the ciliary vibration distinctly in check without determining the death 
of the cell. It is thus possible with solutions of 1-800 to 1-1 GOO to keep 
the cilia moving at a very slow rate, but with quite perfect rhythm, for 
several hours, the movement being often first brought to a standstill by 
the desquamation of the cells. With solutions of from 1-400 to 1-1600, 
the movements may be brought to rest, and then, when desquamation 
has not occurred, restored by irrigation with the salt solution, and this 
may be done not once only, but four or five times. At each successive 
application of the acid, however, the inhibitory effect is more readily and 
quickly produced, until finally all vibrations cease abruptly on contact with 
the acid, and then the salt solution fails to restore it and the cells undergo 
the usual degenerative changes. The spectacle of the gradual re-establish- 
ment of movement on washing out the carbolic solution is extremely inter- 
esting. Sometimes it occurs in a steady progression from one side of the 
specimen to the other, the cilia at first vibrating at their tips through a 
very small arc, and then slowly swaying to and fro, and at last, sometimes 
almost instantly, breaking into a nearly normal rhythmic vibration, with 
the most perfect co-ordination of movement between individual cilia. 
Sometimes the re-establishment of vibration does not occur in a gradual 
uniform progression, but isolated cilia, or groups of cilia commence to 
vibrate independently of their neighbours, and may often be seen in full 
career while those about them are quite still. The same variations may 
be observed in the cessation of movement. After the cilia have been 
brought once or twice to rest by dilute solutions, the amplitude of their 
vibration in the subsequent restorations is, as a rule, much less than nor- 
mal, their rate of vibration slower, and co-ordination of movement in 
neighbouring cilia or cell groups very imperfect. In a few cases it was 
noticed that at the instant of contact of the acid (1-400) with the cells, 
the cilia suddenly stood straight and stiff as if tetanized, and after remain- 
ing so for a few seconds as suddenly broke into rapid vibration, which 
gradually gave place to the slower movement, which is usual under the 
action of the agent. 
The general results of the above experiments may be briefly enumerated 
as follows:— 
Carbolic acid in moderately strong and concentrated solutions (1-100 
and over) causes, either immediately or in a short time, cessation of vibra- 7 
tion in living ciliated cells, with rapid characteristic disintegration of their 
protoplasm and death of the cell. 
In very dilute solutions (1-400 to 1-3200) carbolic acid may cause, if 
long continued, slight degenerative changes in the protoplasm and the death 
of the cell; but its most noteworthy action is its inhibitory effect upon the 
ciliary movement. This may be retarded by it or even entirely checked 
without necessarily determining the death of the cell, for on its replace- 
ment by an indifferent fluid the movement may be perfectly re-established 
and continue indefinitely. 
It is not necessary for the purposes of the present paper to enter into 
the very extensive literature of the ciliary movement, nor to consider at 
length in how far the above described effects of carbolic are similar to 
those produced by a number of other agents, the effects of which, ever 
since the classical investigations of Purkinje and Valentin, have been a 
favorite theme for amateur and professional study. Suffice it to say, that 
most of the agents which bring the vibrations to rest, do so, even in dilute 
solutions, in virtue of their destruction of the life of the cell. To these, 
sulphuric ether, chloroform, and carbonic acid are exceptions. 
It should be further remarked that the power of certain dilute alkalies 
to temporarily restore ciliary movement, which has come to rest either 
spontaneously or by contact with certain fluids, is not analogous to that of 
the salt solution in the above experiments; for the latter acts by simply 
washing out a deleterious by an indifferent fluid, while the former induce 
profound structural changes in the cell, which speedily and invariably 
eventuate in death. 
The Action of Carbolic Action on Living White Blood Cells These 
experiments again fall naturally into two series, in the first of which the 
effect of the acid was studied up in the cells taken from the vessels and 
exposed to its action upon the slide; while in the second its effects were 
observed upon the cells within the tissues and especially upon the phe- 
nomena of emigration. The method employed in the first series was 
essentially similar to that of the first series of experiments on ciliated cells, 
namely, irrigation beneath the cover-glass. Owing, however, to the lia- 
bility of such cells as are fixed upon for observation to be floated out of 
sight by currents of irrigating fluid, great care in manipulation is neces- 
sary, and the following procedure was finally adopted as being the most 
uniformly successful. A small drop of one-half per cent, salt solution being 
placed in the middle of the slide, and the long middle toe of the hind foot 
of the frog1 being carefully cleansed, its tip was snipped off with scissors, 
and the small drop of blood and lymph which presently exudes was care- 
fully deposited in the centre of the drop of salt solution, and covered by 
1 It is very convenient to make use, for this purpose, of frogs, in which an artificial 
leucocythosis has been previously produced by snipping off the apex of the heart and 
allowing considerable blood to flow, and then closing the wound. 8 
thin glass, the cells being protected by thin filter paper on either side. 
The advantage of depositing the blood in the drop of salt solution instead 
of directly upon the slide and mixing afterwards, becomes evident on ex- 
amining the specimen, for under these circumstances the fibrine does not 
form in a close clot, but in a very loose network, frequently exhibiting a 
most exquisite radial arrangement of its fibrillm about the fibrine granules. 
The loose network thus formed, while serving a most admirable purpose 
in preventing the floating otf of the white blood cells, yet interferes but 
little with the free passage of the irrigating fluids in all directions. Irri- 
gation was accomplished by allowing the fluid to drop at slow, regular 
intervals from a fine canula held by a clamp to the stage of the microscope 
and attached to a Mariotte pressure bottle ; the overflow being drawn off 
at the opposite side by a strip of twisted filter paper. 
If a specimen of frog’s blood thus prepared, and irrigated with a half per 
cent, salt solution, be subjected to continuous observation, at a tempera- 
ture favourable to the occurrence of amoeboid movements, these may fre- 
quently be seen to continue for several hours, becoming after a time slower, 
and finally ceasing altogether, as the cells die, when, in a few hours they 
disintegrate, leaving but a few scattered granules behind. If, however, 
instead of irrigating with simple salt solution, a very dilute solution of 
carbolic acid in salt solution (1-800 to 1-1 GOO) be used, the amoeboid 
movements will be found not to continue as long and to be much more 
slowly executed, and to consist often, especially after the first half hour, 
not in locomotor movements, but simply in the extrusion and retraction 
of processes. And, when finally the cells cease to move, they do not dis- 
integrate as rapidly as when the carbolic acid is absent. If the irrigating 
fluid is more strongly impregnated with carbolic acid containing from 
1_800 to 1-400, the movements cease considerably earlier ; while if 1-100 
be employed, they cease almost as soon as the fluid reaches the cell, and 
very soon, if its action be continued, the protoplasm may be observed to 
resolve itself into larger and smaller strongly refractive granules or glo- 
bules and a finely granular translucent substance, which preserves the 
form of the shrunken cell body, the nucleus becoming visible with more 
or less jagged contours. With solutions of 1-50 to 1-20 the movements 
cease instantly, the cells shrink, becoming coarsely granular, and very 
soon exhibit in most cases the above-described disintegration of the pro- 
toplasm.1 
1 The action of carbolic acid on the red blood cells has been very carefully studied 
and described in detail by Huels (Wirkung der Carbolsaure auf rothe Froschblutkor- 
perchen-Inaug. Diss., Anklara, 1872). It will suffice here to say in general, that with 
dilute solutions they shrink, losing their regular contours and becoming variously 
folded and curved, and, after a time, may swell up, become pale, and disintegrate. 
With stronger solutions they become granular, and the protoplasm breaks up, forming 
a homogeneous, brownish-red, strongly refractive material, which may appear either as 
an irregular net-work within the cell or form globules, which tend to coalesce, and 
are insoluble in water, but are dissolved by carbolic acid. 9 
Thus with very dilute solutions of carbolic acid the amoeboid movement 
of white blood cells is held in check, but may still continue for several 
hours. With stronger solutions it either ceases instantly or very soon, and 
the protoplasm undergoes rapid decomposition.1 
Such observations as the above are of course valuable only when made 
in each case with careful control experiments, in which the salt solution 
alone is employed, the other conditions remaining the same; and then 
definite conclusions are justifiable only as the result of a large number of 
observations ; the individuality of the cells in their power of executing 
movements and in their capacity for resisting deleterious agencies, and 
the marked influence of the general well being of the animal from which 
they are taken, upon both, being always borne in mind. 
Carbolic acid enjoys, and certainly deserves, the reputation of being, 
even in very dilute solutions, exceedingly inimical to the life of the lower 
organisms and to cell life in general. The question arises, however, at 
this stage of our investigation, are the cells whose movements have ceased 
through the agency of dilute carbolic acid, really dead ? In the very nu- 
merous and valuable experiments upon its action on infusoria, bacteria, 
spermatozoa, etc., the cessation of movement seems to have been gener- 
ally assumed to be the just criterion of death, and, inasmuch as the con- 
ditions of the experiment are usually so complicated and the organisms so 
minute, it is difficult to prove whether this is justifiable or not, because of 
the difficulty, in most cases the impossibility, of replacing the organisms 
under normal conditions so as to see whether or not their vitality is alto- 
gether lost or only held in abeyance. This, so far as disinfection is con- 
cerned, may, under certain circumstances, be of no little importance, and, 
in the determination of the capacities of protoplasm in general, it is cer- 
tainly of the greatest significance. It has been shown in the first part of 
this paper that carbolic acid in dilute solutions is capable of bringing the 
ciliary movements to rest, and that if then it remains long in contact with 
the cells, they die and disintegrate, but if it is washed away in time, the 
movement may be re-established, showing that in the case of ciliated 
cells, cessation of movement does not necessarily imply the death of the 
cell. Is this true of the white blood cells, brought, as we have seen above, 
into a quiescent condition by dilute solutions of carbolic acid ? If the 
carbolated fluid be carefully washed away from such quiescent cells, before 
it has been very long in contact with them, it will be found, in fact, that 
in a certain number of cases, cells which have been under continuous ob- 
servation, will slowly commence to move again, at first sending out and 
retracting processes only, but at length exhibiting well-defined locomotion. 
1 Essentially the same results were obtained from a study of the action of the acid 
upon the white blood cells of the rabbit and of man. In this case the technique was 
modified by the use of the warm stage, the irrigating fluids being kept at a suitable 
temperature by the water-bath. 10 
But the number of cells which do this is comparatively small. "While 
such results are decisive, as determining that the cells brought to rest by 
means of dilute solutions of carbolic acid are not necessarily dead, it by 
no means follows that those which do not move by a substitution of the 
indifferent for the carbolated fluid are dead, for they are all, at best, placed 
under very abnormal conditions.1 The following series of experiments 
will throw further light upon this subject. 
The action of Carbolic Acid on White Blood Cells within the Living 
Tissues In these experiments the parts used were the tongue, the blad- 
der, and the mesentery of the frog, and the results obtained were essen- 
tially the same for all. 
For the study of the circulation in the bladder and mesentery, modifi- 
cations of the above-described Thoma’s frog plate were used, differing 
from that mainly in the size and shape of the plate upon which the animal 
rests, and the shape and position of the glass upon which the exposed 
part is laid for study. For the observation of the normal circulation and 
the phenomena of emigration the bladder is unsurpassed among all the parts 
of the frog which are used for this purpose. To prepare this organ for study, 
a lateral incision is made in the abdominal wall of a curarized animal, 
most conveniently on the right side, extending from the posterior end of 
the trunk about half way to the axilla, care being taken to avoid or ligate a 
large cutaneous vein which usually runs near the line of incision. A bent 
canula attached to a large pipette, filled with a half per cent, solution of salt 
is then introduced into the cloaca and directed forward into the bladder, 
and fastened in position with a’bit of thread passed through the skin just 
above the anus. The salt solution now being injected into the bladder, 
either by the breath or by attachment to a suitable pressure flask, the 
organ as it fills is crowded out through the incision, and, being held in 
distension by a spring clip attached to the distal extremity of the pipette, 
it is laid upon the glass disk provided for that purpose in the frog 
plate and may be covered with thin glass and irrigated in the usual way. 
It can be examined with high powers, and leaves little to be desired in 
the clearness and precision with which both the normal and pathological 
conditions may be studied. It is, however, less well adapted for long 
continued observations of a special limited area or of a single cell, than the 
mesentery or tongue, because of the slow movement of the organ, due to 
the contraction of the smooth muscle cells so abundantly interlacing in its 
walls. This movement is more troublesome under ordinary circumstances 
than that induced in the mesentery by the intestinal contractions, because 
1 Special stress is laid upon this point, not because the capacity of reducing the 
activity of amoeboid movements or bringing them to rest without killing the cells, is 
unique in carbolic acid, for we are already acquainted with many other substances 
which possess the same capacity, such as strong salt solutions, quinia, etc.; but for 
the sake of emphasizing in reference to cell activity, its inhibitory in distinction from 
its destructive power. 11 
the bladder is exposed to the air over a greater area, and is less readily 
confined. It is worthy of note, however, that carbolic acid, even in solu- 
tions so dilute as to interfere but little with the circulation, very soon 
reduces this contractility of the smooth muscle cells of the bladder to such 
nearly complete inactivity, that the observation is but little interfered with. 
The methods of preparing the mesentery and tongue for study are so well 
known that nothing need be said liere about the technique. 
In all of these experiments, for obvious reasons, as small a quantity of 
curare was used at a dose as was consistent with the attainment of suffi- 
cient immobility of the animal. 
If the freshly exposed bladder or mesentery of the frog, in which the 
circulation is active, be irrigated for an instant with a strong solution of 
carbolic acid (1-20) immediate stasis is produced in all but the larger 
vessels, the part becomes cloudy, and both white and red blood cells be- 
come rapidly shrunken and distorted, and degenerate in the manner above 
described, when the effect of the same solution was studied upon the ex- 
travasated cells. The bloodvessels change their calibre but little before 
stasis occurs in the larger trunks also, and the part very soon shrivels and 
commences to reveal the retrogressive metamorphoses which an entire 
absence of fresh blood involves. If these parts be irrigated with a more 
dilute solution (1-50 to 1—100) they soon become somewhat cloudy, and 
stasis will be seen to occur almost immediately in many of the capillaries 
(except those which form direct communication between larger arterial 
and venous trunks), gradually extending to the smaller and larger veins, 
and then to the arteries, which often dilate considerably, but without, as 
a rule, any perceptible acceleration of the current. One after another 
these vessels become crowded with red and white blood cells, the plasma 
becoming less and less in quantity, and usually within from twenty to 
ninety minutes all movement ceases, except in the largest trunks, in which 
a slow current may often continue for a considerable time longer. The 
details of the manner in which this stasis occurs may be best observed in 
the mesentery with solutions of 1-100, and with especial clearness in the 
smaller veins. One may observe in these, as the process advances along 
them from the direction of the capillaries, that the red blood cells no 
longer all present a symmetrical form, but are variously distorted, perma- 
nently curved or bent, sometimes appearing slightly swollen, while here 
and there one or more may be seen sticking closely with its side against 
the wall of the vessel. To this • others may join themselves, forming a 
little heap of distorted cells encroaching upon the lumen of the tube, or, 
instead of being piled up in heaps, the red blood cells may lie along the 
wall, side by side, in a single or double layer, thus often forming a com- 
plete cylindrical investment to the inside of the vessel, through which the 
blood current may still pass with considerable, though markedly impaired, 
velocity. Gradually, however, more and more accumulation occurs until 12 
the lumen of the vessel is finally entirely occluded, and the current stops. 
Then one may see in many cases how the cells become crowded closer and 
closer together until at last a uniform red mass fills the vessel, which soon 
becomes partially decolorized, and shows the nuclei with great distinct- 
ness. Sometimes the little collection of cells formed along the walls are 
loosened by the current, and driven along out of sight into the larger 
channels. 
We have thus a most exquisite example of the formation of thrombi 
and emboli caused, partly at least, by evident changes in the red blood 
cells of which they largely consist. The white blood cells, under these 
circumstances, do not tend to collect in the peripheral zone of the current, 
which is very little developed, however long the circulation may persist; 
they invariably retain the spherical form, and are mingled indiscrimi- 
nately with the red cells, and when the latter come to rest remain entan- 
gled among them, forming a small part of the thrombotic mass. This 
condition of affairs, already described by Ilueter,1 under the name of 
“ globular stasis,” although, according to that observer, readily undergoing 
resolution in the skin of the frog, where he chiefly studied it, does not do 
so when far advanced, even under the most favourable circumstances, in 
the bladder and mesentery. The appearance of the red blood cells as 
they lie against the walls, under these conditions, is not at all to be con- 
founded with that often seen in diapedesis of the red blood cells. Where 
the latter process is occurring one or many cells may, indeed, be seen 
clinging to the wall of the vessel, but that part of the cell which is within, 
if it form a considerable portion of it, is pendulous, adhering at one point, 
and waves to and fro in the current, and is beaten about by passing cells. 
Nothing of this kind is seen in the cells changed by carbolic acid ; they 
almost invariably lie flat against the wall, seem to be stiff and unyielding, 
and appear to have entirely lost their elasticity. That these effects are 
due to a purely local cause, is well shown by the fact that the distortion of 
the red cells and their adherence to the walls, and to one another, is first 
seen, arid most evident throughout, either in the thinnest walled vessels, 
or in those in which the circulation is comparatively slow. Where fresh 
blood from the heart is being rapidly driven through the vessels, on the 
other hand, even though they be quite narrow and thin walled, as in some 
of the capillaries which form a short or direct communication between 
large arteries and veins, the changes are tardy in appearing. 
If we turn now from solutions of carb.olic acid of such concentration as 
to cause permanent stasis within a short time in these delicate parts, to 
those of such dilution that under their action the circulation may still con- 
1 Hueter, Algemeine Chirurgie, §§ 193 and 235 ; also Deutsche Zeitsclirift fur Chi- 
rurgie, Bd. 4, Heft 2, u. 3. This condition may, according to this writer, be produced 
by a number of agents besides carbolic acid, among which are alcohol, ether, chloro- 
form, ammonia, glycerine, etc. 13 
tinue active for several hours, namely, to solutions of the strength of from 
1_800 to 1-3200, we find that within that range of dilution the effects 
produced are closely similar, and that when the parts are carefully pre- 
pared the circulation may often continue, with no tendency to stasis or 
thrombosis, for at least twenty-five hours, beyond which time observations 
were not extended. For the sake of convenience we may confine our 
attention to the effects produced by a solution of the strength of 1-1 GOO, 
which may be regarded as typical. If we expose either the bladder or 
mesentery, and irrigate for a few moments with one-half per cent, salt solu- 
tion, until the animal has recovered from the shock of the operation and 
the circulation has adapted itself to its new conditions, and then, cutting off 
the salt, irrigate with the carbolated solution of the above strength, we 
very soon observe a slight dilatation of both veins and arteries, with no 
marked change in the capillaries. This dilatation rarely exceeds one- 
eighth to one-quarter the original calibre of the vessel, and is permanent 
as long as the part is exposed to the carbolated fluid. The blood current 
becomes at the same time slightly accelerated for a few moments, and then 
usually becomes slower than normal, and so continues. At first the usual 
distribution of blood in the veins and arteries may be noticed, the axial 
current and the peripheral plasma zone, and within the latter, in the veins, 
a greater or less number of white blood cells may be seen rolling or glid- 
ing slowly along the walls, and even permanently adherent to them. Very 
soon, however, the plasma zone becomes less distinct, while the axial zone 
becomes somewhat broader, and by little and little a large proportion of 
the white blood cells which were at first fixed to the walls become loosened, 
and are carried off by the current.. AVhile still a considerable number are 
seen rolling along in the periphery, and even exhibit to a certain degree 
the well-known appearance of stickiness, this is very much less marked 
than when an indifferent fluid is used for irrigation, and very few cells, 
indeed, become permanently fixed. Those, moreover, which do drag 
along in the peripheral zone are, for the most part, globular, and are not 
drawn out into the well-known pyriform shapes presented under ordinary 
conditions. This condition persists as long as the carbolated fluid is con- 
tinuously applied, and thus twenty-four hours and more of almost unre- 
mitting observation may pass, without the detection of a single cell in the 
act of passing through the walls, or moving in the tissues outside. Such 
experiments were often repeated, and while in no case was a cell actually 
seen passing through the wall, a few times, a small number of cells, having 
the characters of white blood cells, were seen to have appeared in the tis- 
sues outside the vessels, but in no case at a great distance from them. 
We are thus lead to the conclusion that, under the above conditions the 
irrigation of the mesentery or bladder of the frog with very dilute solu- 
tions of carbolic acid (1-1 GOO) prevents, almost entirely, the emigration 
of white blood cells. 14 
If now the carbolated fluid be replaced by one-lmlf per cent, salt solu- 
tion, the picture very soon changes. In the course of two or three hours, 
often much sooner in the bladder, without any marked change in the 
calibre of the vessels, the white blood cells begin to collect in the peri- 
pheral zone of the veins. They seem to be much more adherent to the 
walls as they glide along; they are seen to catch now and then at some 
point, and become elongated and drawn out into pyriform shapes, and very 
soon a greater or less number may be seen to be permanently fixed in the 
walls, through which certain of them are readily seen to slowly pass and 
wander oflf in the tissues. The process of emigration, then, may be held 
in check by dilute solutions of carbolic acid, but on the replacement of the 
latter by an indifferent fluid it readily occurs. If now, with emigration 
in full course, the carbolated fluid be again applied, a few cells may still 
be observed passing out of the vessels at first, but their movements soon 
become extremely slow, and shortly cease altogether; most of the white 
blood-cells in the peripheral zone gradually assume the spherical form and 
become less adherent, and the plasma zone is finally, to a considerable 
degree, freed from them. 
Thus three times, in the same animal, in the course of twenty-four 
hours the alternation of irrigating fluids was accomplished, and each time 
with the same result. 
These experiments may be varied in a number of ways. Thus the 
mesentery may be simply exposed to the air under irrigation with one-half 
per cent, salt solution, until emigration is actively progressing and the 
cells moving through the tissues and coming to the surface between the 
endothelium in the manner described in detail by J. Arnold,1 when, on 
application of the carbolated fluid, not only does emigration soon cease, 
but also the locomotion in the tissues, and cells which were working their 
way slowly through on to the surface of the endothelial investment are 
brought to rest and so remain, often with a little shining knob through 
on the outside, while the remainder may be more dimly seen perfectly 
quiescent beneath. Cells which had been under observation in this con- 
dition for several hours were seen, on the replacement of the carbolated 
by the salt solution, to slowly drag themselves through on to the surface 
where they were floated off by the irrigating fluid. Still again, if a wound 
be made in the tongue by carefully dissecting off a bit of the mucous mem- 
brane from the papillary surface, emigration may be almost entirely pre- 
vented for many hours by irrigating with the dilute carbolated solution ; 
or, if allowed to progress under an indifferent fluid, it may be suppressed 
at any time in the same way. 
The most careful scrutiny failed to reveal any structural changes in the 
1 Arnold, Ueber die Durchtrittstellen der Wanderzellen durch entzundete Seiose 
Haute, Virch. Archiv, Bd. 74, p. 245. 15 
white blood-cells rendered quiescent within the vessels or the tissues, by 
such dilute solutions as 1-1600. 
These experiments were performed exclusively upon spring and sum- 
mer frogs, which were for the most part freshly captured. Large or 
vigorous animals were invariably chosen, and, as above stated, a minimal 
dose of curare was always administered. Nevertheless it is not surprising, 
considering that the lethal dose for the frog is so small,1 and that large 
quantities of the irrigating fluid were used to ensure uniformity in the 
conditions of the experiment, that in a certain small number of cases, 
when the irrigation was long-continued, a sufficient quantity was absorbed 
to cause death, or at least a considerable feebleness of the heart’s action 
before the conclusion of the experiment.2 All such cases were, of course, 
excluded in making the summary of the experiments on the action of the 
acid. 
Conclusions The chief results of these investigations are as follows:— 
Strong solutions of carbolic acid cause in living, active, ciliated, and 
white blood-cells, immediate cessation of movement and death of the cells, 
with speedy disintegration of the protoplasm. 
Very dilute solutions of carbolic acid cause retardation or temporary 
cessation of movement in living, active, ciliated, and white blood cells, 
which may, under suitable conditions, be restored by the replacement of 
the carbolated by an indifferent fluid. 
Strong solutions of carbolic acid, if applied to the bladder, tongue, or 
mesentery of the living frog, speedily cause stasis and thrombosis, partly, 
at least, through profound changes induced in the protoplasm of the red 
and, perhaps, the white blood cells. 
Very dilute solutions of carbolic acid, if applied to the bladder, tongue, 
or mesentery of the living frog, under conditions in which inflammatory 
changes commonly occur, modifies the usual character of these, by pre- 
venting the occurrence of any considerable degree of emigration or loco- 
motion of white blood cells in the tissues, the usual condition of affairs 
supervening, however, if the carbolated be replaced by an indifferent 
fluid. 
One word should be added in conclusion, in reference to the relation of 
the above results to the process of emigration in general, and to the heal- 
ing of wounds under the action of carbolic acid. 
1 Plugge, Pfliiger’s Arch., 1872, Bd. v. p. 5-10, estimates it as 0.010 grm. 
a It must be confessed that this assumed relation between the feebleness of the 
heart’s action and the absorption of the acid is, in large measure, conjectural; for, 
in addition to the incompleteness of our knowledge concerning the precise action of 
carbolic acid.upon the heart under ordinary circumstances, the condition of the animal 
here, under the influence of curare, would doubtless render the exact condition of 
affairs still more obscure. 16 
First. The more intimately we become acquainted with the process of 
emigration the less does it seem to possess the comparatively simple char- 
acter with which it was at first credited. Neither the physical conditions 
determined by pressure, nor the relative or absolute position of the white 
and red blood cells within the vessels, nor the assumed glutinosity of the 
former, nor the existence of minute currents of fluid passing under cer- 
tain conditions through the walls of the vessels into the lymph spaces out- 
side, nor, finally, the locomotor capacities of the white blood cells them- 
selves, seem alone to sufficiently explain this very curious and complex 
phenomenon. It seems probable that not the influence of one of them 
alone, but of all combined, and perhaps of others of whose existence we 
are not yet aware, is essential to its accomplishment. If this be true, we 
are at present by no means sufficiently acquainted with the action of the 
several factors to assign to each its appropriate share in the accomplished 
result. The latest supplement to the story of the investigation of the ac- 
tion of quinia on emigration1 teaches, that it is by no means improbable 
that its inhibitory action is, in some cases, at least due to the change of phy- 
sical conditions which it induces in the circulating blood, and not entirely 
to its power of restraining protoplasmatic movements. 
Moderately concentrated solutions of salt also possess the power, as 
shown by Thoma,* of checking emigration, largely, probably, by the in- 
duction of a simple change in the physical conditions of the cells, and a 
dilatation of the bloodvessels and a consequent increase in the rapidity of 
the current. So in the above experiments, while the carbolic acid un- 
questionably exerts a paralyzing action upon the white blood cells, and 
seems to have little evident effect, so far as microscopical observation can 
determine, upon the bloodvessels, we can still only say with safety at 
present, that its action upon the white blood cells is at least one of the 
ways in which it holds emigration in check. 
Finally, as to the relation of the results of these experiments to the use 
of carbolic acid in the treatment of wounds. One of the prominent claims 
to virtue in its action, both by the devotees to Listerism and the advocates 
of a modified use of the drug is, that it restrains undue suppuration. 
Unfortunately the writer had not at his disposal the facilities for studying 
the action of carbolic acid upon the process of emigration in the warm- 
blooded animals, and the value of such observations upon the frog, as those 
described above, when applied to warm-blooded animals, and to man, each 
reader must finally decide for himself. The limitations, however, within 
which observations on one class of animals may be safely applied to an- 
other, are becoming daily more clearly defined, and it would seem to the 
1 Appert, Der Einfluss des Chinin auf die Auswanderung der weissen Blutkorper 
bei der Entziindung. Virch. Archiv, Bd. 71, p. 364. 
* Thoma, Der Einfluss der Concentration des Blutes u. d. Gewebssafte auf d. Form, 
u. Ortsveranderung farbloser Blutkorper. Virch. Archiv, Bd. 64, p. 1. 17 
writer, at least highly probable, judging from the above experiments, that 
a part of the favourable action of carbolic acid in restraining undue sup- 
puration in wounds is due to its direct effect upon the process of emigra- 
tion, in so far at least as pus is the result of this process, and not of the 
proliferation of other cells. Whether beyond this it may or may not act 
beneficently in killing micro-organisms, or whether it may or may not 
have other less well-defined beneficial effects upon the healing process, are 
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