Technical Report No. IRL 1110

CYTOCHEMICAL STUDIES OF PLANETARY MICROORGANISMS
EXPLORATIONS IN EXOBIOLOGY

Summary Report Covering Period July 1, 1969, to July 1, 1970
For
National Aeronautics and Space Administration

Grant NGR-05-020-004.

 

Instrumentation Research Laboratory, Department of Genetics
Stanford University School of Medicine
Stanford, California 94305
Report to the National Aeronautics and Space Administration

"Cytochemical Studies of Planetary Microorganisms - Explorations in Exobiology"

NGR 05-020-004

Summary Report Covering Period July 1, 1969 to July 1, 1970

Instrumentation Research Laboratory, Department of Genetics
Stanford University School of Medicine

Stanford, California

 

hua Lederberg
rincipal Investigator

Elliott C. Levinthal, Director
Instrumentation Research Laboratory
A. INTRODUCTION

This report covers the activities of the
Instrumentation Research Laboratory during the two
quarter period from January 1, 1970 to July 1, 1970 and
serves also as a summary annual report of our work for

the year ending July 1, 1970.

While the main support of the [IRL activities during
this period continued to be the NASA grant
NGR-05~-020-004, some of the funds have come from other
grants, other agencies, and in some cases private
Institutions. This report includes all the activities
of the laboratory which relate to or have benefited
from NASA support regardless of whether or not. they

were primarily supported by this NASA ‘grant.

We have applied for funding for our work from. other
agencies, particularly the National Institutes of
Health, with some degree of success, especially in the
area of cell separation. Effective January 1, 1970
essentially all of our work in this area was supported
by NIH grant GM 17367 and NIH Contract 69-2064. We have

received some support for Dr. Halpern's work under NIH
Grant AM 12797 and are seeking additional support for
his work on the interaction of chlorine compounds with
DNA. Our work on computer aided instrumentation has
been partially supported by a collaborative program
with the Physics Department and their Air Force
contract, AF F 44620 67C 0070. The efforts on image
processing for the 1971 Mars Mariner are a joint effort
with the Artificial Intelligence Laboratory of the
Computer Science Department under Professor John
McCarthy, Advanced Research Projects Agency grant SD

183

These Interrelationships benefit our NASA program in
two ways. They ald the rapid utilization for medicine,
and blology in general, of the ideas and skills
developed because of our interests In space missions.
They not only provide us with the opportunity of
testing instrumentation methods, applicable to future
NASA programs, in circumstances of solving current
scientific problems but also provides much needed
additional support. This support hopefully will permit
the Instrumentation Research Laboratory to continue to
function with sufficient technological depth and

breadth to allow responses to future NASA needs.
For these reasons we have felt it desirable to carry
out some laboratory work that could lead to new
fruitful relationships. A meeting on Space Bloscience -
Technology Utilization held on November 28-30, 1969
under the auspices of the Interdisciplinary
Communication Program, was attended by Professor
Lederberg and Dr. Levinthal. This meeting reinforced
our conviction that there are areas of environmental
health that would be an intellectually challenging and
socially productive use of our scientific and
technological skills. During this reporting period we
have begun to _ investigate such possibilities in
environmental health, particularly in environmental

monitoring.
The general areas of the program resume, Part B of

report, are:
Topi

It.
Iti.
IV.

Vv.

VI.

Vil.
Vill.

1X.

Xt.
Xt.

Xltt.

c

The Determination of Optical Activity
by Gas Liquid Chromatography

Gas Chromatography of Amino Acids

The Reaction of Chlorine with DNA

Apollo 11 and 12 Sample Analysis

The Mass Spectrometry of
(l-phenylethy!)-Carbamates

Analysts of Natural Products by
Mass Spectrometry

DENDRAL

Computer Aided Instrumentation

Cell Separation

Optical Data Processing

Quasi Microscope for Mars Lander

Mariner Mars 1971 Orbiting Photography

Reports, Publications and Papers

the

Page

10

11
13
15
20
25
32
34
36
B. PROGRAM RESUME

1. The Determination of Optical Activity by

Gas Liquid Chromatography

As members of the team which is interested in the determination
of optical activity of amino acids in planetary soils, we have
collaborated with Ames personnel in the construction of a working
model of a hydrolyser-desalter-derivatizer unit. A miniaturized
version of such a unit will be compatible with the

GC-MS-pyrolyser experiment proposed for the 1975 Viking mission.

Our chemical work on the GLC separation of diastereoisomers' has
been continued. Useful applications have been in the
determination of optical purity of. alcohols, ketones,
phenylacetic acids and phenoxypropionic acids. This analytical
technique has also been used _ to determine the absolute
configuration of alloisoleucine present in the serum of a patient

suffering from "Maple Syrup Urine disease".
It. Gas Chromatography of Amino Acids

We have developed a simple technique, which permits the gas
chromatography of some amino acids, without time consuming
chemical manipulations. The procedure uses the injector port of a
gas chromatograph as a chemical reactor. Since these conversions
are concentration dependent, the injection port of a conventional
chromatograph had to be modified to permit the removal of the
solvent before the pyrolysis reaction. A prototype of a partially
automated sample injection system has been constructed and the
new procedure has been used to determine blood phenylalanines. A
completely automated version of such an instrument should be of
use in the routine screening of newborns for cases of

phenylketonuria.
fil. The Reaction of Chlorine with DNA

Chlorine plays an indispensable role in the purification of our
water supplies and urban settlement would be imperiled without
chlorination. Despite our great dependence on the _ chlorine
disinfection of water, the potential chronic toxicity to man from
chlorine has not been considered a hazard, because chlorine is
known to react rapidly with organics, which would lead to
destruction of the reagent In the body fluids. This conclusion
however ignores the possibility that the chlorine reacts with a
variety of nitrogen containing compounds to form chloramines,
which may themselves form potent chlorinating agents. Whilst
there is an extensive literature on the stable end products of
chlorination of the purines and pyrimidine bases and _ their
nucleoside derivatives, chloramine derivatives of these compounds

have not been described.

We have now examined the chlorination products obtained from
cytosine and cytidine and find that they behave as_ typical
chloramine derivatives. The compounds give a test for positive
chlorine (Von Arx-Neher) and liberate fiodine from Ki solution. We
have used the latter reaction to quantitatively estimate’ the
amount of labile chlorine in the products. Under essentially the
same experimental conditions and using between 0.01 to 10.0
equivalents of chlorox per base of DNA, we find that there is

little change in the molecular weight of the "modified" DNA as
determined by sucrose gradient or elution on Sephadex 200. A
large excess of chlorox leads to breakages of the DNA and the use
of very concentrated solution of DNA results in the formation of

some very high M.W. "modified" DNA.
IV. Apollo 11 and 12 Sample Analysis

Under Contract NAS 9-9439, we have examined the carbon compounds
present in two samples of Apollo 11 and two samples of Apollo 12
for "porphyrin-like" pigments. Evidence for such compounds’ was
obtained in Apollo 11 and 12 IJunar fines. The pigments’ from
samples collected near the lunar lander were’ probably due to
contamination by rocket exhaust products. A porphyrin-like
pigment (+5 x 107>. g/g) was also found in an Apollo 12. sample
which had been collected at a point well removed from the’ Junar

landing site.
-10-

V. The Mass Spectrometry of (1-Phenylethyl)-Carbamates

This class of organic compound was used for the optical analysis
of asymmetric secondary alcohols. During this work several
interesting features were noted in the mass spectra of these
carbamates. In oorder to investigate the precise fragmentation
modes of these molecules several deuterated compounds were
synthesized. After studying the mass spectra of the carbamates
labeled with deuterium it was possible to formulate
rationalizations for the principal fragmentation processes
including those hydrogen atoms transferred from one portion of
the molecule to another. To avoid thermal decomposition it was
necessary to use the direct inlet system and to maintain the ion
source at temperatures below 160° or extensive thermal

fragmentation of the carbamates resulted.
-~ll-

VI. Analysis of Natural Products by Mass Spectrometer

During the past year research has continued on the structural
analysis by mass spectrometry of natural products isolated
from plant, animal and marine sources. A list of publications

resulting from this experimentation follows:

1. "Mass Spectrometry in Structural and Stereochemical
Problems - CLXXII* The Electron-Impact Prompted
Fragmentation of 1,2-Cyclohexanediol" by M. Karen
Strong and Carl Djerassi. Org. Mass Spectrometry
2, 631 (1969).

2. "Application of lon Cyclotron Resonance to the
Structure Elucidation of the CHE OT fon Formed
in the Double McLafferty Rearrangement" by
Georsze Eadon, John Diekman, Carl Djerassi.

v. Am. Chem. Soc. 91, 3986 (1969)

3. "Mass Spectrometry in Structural and Stereochemical
Problems CLXXVI. The Course of the Flectron Impact
Induced Fragmentation of Androstane" by L. Tokes
and Carl Djerassi. J. Am. Chem. Soc. 91, 5017 (1969).

4. "Mass Spectrometry in Structural and Stereochemical
Problems CLXXVI!II. The Electron-Impact Promoted
Frasmentation of 1,2-Cyclohexene Oxide" by M. K,
Strong, P. Brown, C. Djerassi: Org. Mass

Spectrometry 2, 1201 (1969).

5. "Mass Spectrometry fn Structural and Stereochemical
Problems CLXXIX. The Electron Impact Induced
Rearrangements of [-Phenylheptenes. Further Evidence

for Double Bond Lability" by A. F. Gerrard and
C. Djerassi. J. Am. Chem. Soc. 91, 6808 (1969).

6. "Mass Snectrometry in Structural and Stereochemical
Problems. CLXXXIIT!. A Study of the Electron Impact
Induced Fragmentation of Aliphatic Aldehydes" by
R. J. Liedtke and C. Djerassi. J. Am. Chem. Soc.
91, 6814 (1969)
-12-

"Mass Spectrometry in Structural and Stereochemical
Problems CLXXXI!. tnvestigations in the 10-Phenyl-
2-decalone System. The Synthesis and Electron
Impact Promoted Phenyl Migration of trans-10-Phenyl-
~2-octalone" by R. T. Gray and C. Djerassi.

J. Org. Chem. 35, 7533 (1970).

"Mass Spectrometry in Structural and Stereochemical
Problems CLXXX!. Further Studies of Remote Group
Interactions After Electron Impact in 4-Substituted
Cyclohexanones" by R. T. Gray, R. J. Spangler and
Carl Djerassi, wY. Org. Chem. 35, 1525 (1970).

"Mass Spectrometry in Structural and Stereochemical
Problems CLXXXIV. Charge Localization in Fraement
lons of Amino Ketones and Fsters." J. Cable, G. W.
Adelstein. Org. Mass Spectrometry 3, 439 (1970).
-13-

VII. DENDRAL

During the past year, we have essentially completed phase 2 of
the Dendral project, the demonstration of the utility of
artificial intelligence as an aid in the solution of problems’ In
analytical organic chemistry. (Phase 1, 1964-1967, was the system
definition and general analysis of the problem of organic
structures; Phase 2, 1968-70, has been the production and
debugging of working computer programs.) A serles of
publications, some still in press, have exhibited the application
of these methods to different classes of non-cyclic molecules.
Ref. 32 also illustrates the extension of the program to some
simpler cyclics and shows the feasibility of further extension.
Ref. 33 is a general overview of the work to date. The procedures
are worked out for data from mass-spectrometry, infra-red and NMR
instrumentation.

We have now begun work on further phases.

Phase 3: "Meta-Dendral", a program whose output would be = an
encylopedia of chemical reaction rules for the behavior of
compounds in the mass spectrometer. Its Input would be’ empirical
data from compounds of known structure. This work is more more
revealing than DENDRAL itself is of the thought processes used by
the chemist in inducing generalizations. It should also give a
more reliable, self-consistent theory of mass spectrometry than
is now available to DENDRAL from its human instructors. Fragments

of the program have been written so far.
-1lh-

Phase 4&: "Synthetic Dendral", a program to design and
theoretically test reaction sequences for the synthesis of
specific, destred end products. This has already been

demonstrated in a preliminary running version.
-15-

Vill. Computer Aided Research Instrumentation

The work in computer instrumentation of laboratory Instruments,
principally mass spectrometers, is undertaken’ primarily to
investigate techniques applicable to exoblology missions. It
supports much of the work described in this report and in
addition has had application to several laboratories of this and
other institutions, as well as civilian technology spin-off. The
computer Instrumentation has made use of ACME (Advanced Computer
for Medical Research, a Stanford Medical School computer
development utilizing an 1BM 360/50) with an [BM 1800 to serve
individual laboratory instrumentation tasks in a time shared
mode. ACME is primarily supported by NIH grants, but here again a
deliberate policy is followed to merge goals and grant resources
with other research groups at Stanford to optimize joint
objectives. These instrumentation developments have had notice
and value beyond their use here at Stanford. They have been well
reported in the scientific literature, the trade journals, and
seminar papers. (Status reports have included bibliographies
covering each reporting period.) Among these developments are a
unique mass spectrometer control system (which now {s_ available
as aocommercial product), mass spectrometer data processing
systems, and a spectrophotometer data acquisition and processing

system.

Much of our effort of scientific communication has been In
-16-

connection with the more general aspects of our work, namely the
asynchronous time-shared use of a computer for stmultaneous” real

time instrumentation in several different laboratorles.

A TIME SHARED COMPUTER INSTRUMENTED LABORATORY

 

MASS SPECTROMETER
OR OTHER INSTRUMENTS
WITH ELECTRONIC OUTPUT

 

 

 

  
 

 

 

 

 

 

 

 

 

 

 

SPECIAL PURPOSE INTERFACE | Digr TAL
That
| || [LABORATORY
i | INSTRUMENTS
a mit COMPUTER
SYSTEM
¢ til
26
AKALOG OATA OIGITAL DATA
a Lf
6 AND/OR RND/OR a
1 OUTPUT ourpuT {f_____/
COMPUTER DATA TIE PANEL TERMINAL

 

 

 

 

Lo A SYSTEM (PROGRAM) >+—_—_
Ls THE TIME SHARED COMPUTER |

 

Figure 1. Typical laboratory instrumentation center contains computer
data tie panel, keyboard terminal and graphic plotter.

Except for the special development, reported during the first
half of this year, to support the lunar investigation of
porphyrins by magnetic circular dichroism spectroscopy, this year
has been characterized by the use and operation of computer
systems started in prior reporting periods. A critical review of
these projects has led to the definition of a new development we
have now undertaken, the HIQ (High Intelligence Quotient)

Terminal. This is described in more detail later In this section.
-l17-

Figure 1 represents the basic features of a typical time-shared
computer instrumentation problem. The specific design of these
features affects the developmental cost and the degree of
sophistication attainable. The choice of how much of the data
channel (extending into the Computer Data Tie Panel in Figure

1 should be part of the computer system or the special
purpose Interface, the type of signal conditioning available to
the user, the program language, and other variables all influence
the level of computer assistance achleved by computer-managed

instrumentation and its cost.

Over the last several years we have gained valuable experience in
a variety of systems, ranging from a small stand-alone laboratory
computer, such as the LINC, to a bare terminal connected to a
remote large time-shared computer (ACME). This experience coupled
with the development of less expensive "mini computers", has led
to the concept of the "smart" terminal that provides local
control for the instrument in the laboratory. This “terminal"
with enough computer power to process the data stream allows a
connection, using phone lines, to a larger computer system for
maximum programming power, complex data manipulation, and large
data~base filing. Our interest developed in a general modular
design of such a terminal. Professor Melvin Schwartz of the
Physics Department was led to a similar conclusion based on his

instrumentation needs at SLAC (Stanford Linear Accelerator
-18-

HIQ Time shared computer TERMINAL

using DIGITAL PATHWAYS modules

 

 

 

pdpil_—

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

3" ROCESSING _
| (CPU
| UNIBUS
le Ls! 8,
S08 y 00; 00:
E Dg) 8 700: 00:
=8 n= oo oo
° ?

 

 

 

PLUG-IN MODULES INCLUDE DIGITAL INPUT,
DIGITAL OUTPUT, A-to-D, D-to-A PLUS OTHERS

 

TELEPHONE
MODEM

 

 

TO THE TIME
SHARED COMPUTER

SS FACILITY

 

 

 

 

AUXILIARY
KEYBOARD

fj

 

 

 

 

v
*

 

 

UNIBUS EXTENSION

 

 

ESPECIALLY SUITED TO LABORATORY SUPPORT OF MEDICAL,
CLINICAL, BIOCHEMICAL,AND PHYSICS APPLICATIONS.

SOME UNIQUE PLUG-IN MODULES ARE:

 

 

 

 

 

 

 

EXTENDABLE 3) fi
MEMORY : Aa
IN2 OR 4K i() Te
INCREMENTS 50 Mhz sUcvack

HISTOGRAM scapes 20 u:

DISPLAYS 0°

ns "6 TIME BASE
0 Obes TAPE 2 RAPHI QpciLo ep CONTROL
: CASSETTE ZTERMIN : 7 NTR
oh HOLLER FOB HARD :OBbut {S| p— ANALOG
TO: 2) GQPY SRT
we ‘Alotyers OH LAY RECEIVERS
O° °° uTPuT OSCILLOSCOPE oo

 

 

 

 

 

 

 

Figure 2

THE UNIBUS ALLOWS DATA
TRANSFERS BETWEEN
THE LPU AND ANY MODULE.

IN ADDITION, WITH THE “BIN
CONTROLLER”, DATA
TRANSFERS MAY BE MADE
BETWEEN MODULES EVEN
WITHOUT THE LPU

 

 

 

 

 

 

 

 

 

 
  
 

CABLES
FOR ANALOG

DATA ¥ DISPLAY
AND ACQUISITION
-19-

Center) as have other students of NASA's general data-handling
requirements. They had an advanced modular concept locally termed
"Digital Pathways". Together with Professor Schwartz we have
started an interdisciplinary, interdepartmental project to
develop what we have termed the "HIQ" terminal with three groups
of needs in mind, biochemical research, physical research, and
clinical laboratory. This is supported jointly by our NASA grant

and AF contract AF F 44620 67C 0070.

Figure 2 is a_ functional diagram of this modular system. The
present funds allow the development of a pilot model. These will
consist of the PDP-11 and the direct memory access (DMA) design
that is basic to most, if not all, the modules. The DMA portion
is to be the left half as shown with connectors and indicator
lights. Present plans are to built three or four complete module
types to connect to the PDP-11 Unibus. This initial unit will be
used to improve service on the MS-9 mass spectrometer in the

Organic Chemistry laboratories at Stanford.

After this initial experience it is hoped to build additional of
the functional modules and construct an uncommitted system for
experimental use. [It will be a new type of instrument in that [It
will bring the computer to the test and research environment as a

tool and not a project in itself.

As part of this pilot development we expect to be able to
estimate the production cost of such terminals. We believe’ that

they will prove to be economically practicable.
-20-

IX. Cell Separation

While this work was initiated by the subject grant most of the
support is now coming from NIH grant (NIH GM 17367) and NIH
contract (NIH 69 2064). The work has obvious applications In the

medical field.

A. High Speed Fluorescent Cell Sorter

This unit is designed to measure the fluorescence of cells In a
jet of liquid, break up the jet into uniform drops and collect
the drops in a sertes of containers, with all drops containing
cells with similar fluorescent characteristics collected In the
same container. Tests showing that the system could enrich for
plaque forming cells, using the Jerne plaquing technique have
been reported in Science. A second unit using a sheath flow
system, which encloses the stream of fluid containing the cells
in a larger stream of inert liquid has been built and tested.
This unit possesses superior optical properties and appears to be
more sensitive than the ortginal. A blue laser, now on order,

should increase the sensitivity even further.

B. High Speed Volumetric Cell Sorter
This unit measures the volume of cells by determining the change
in electrical resistance In a small orlfice containing flowing

conducting liquid as the essentially nonconducting cells pass
-2?1-

through. The sheath flow principle has also been applied to this
instrument. Downstream of the orifice a jet can be formed = and

deflected in the same fashfon as In the fluorescent sorter.

Measurement of the volume distribution of red blood cells in this
instrument indicates that sheath flow gives a more accurate
reproduction of the expected approximately Gaussian volume
distribution than measurements where the entire orifice is
avallable to the cells. This is indicated in Figure 3 where curve
A shows the red cell volume distribution In sheath flow, compared
to curve B showing a run using the same orifice when the outer
sheath is no longer flowing. Curve C shows the response to. the
same blood sample of a commercial instrument for measuring red
cell volume which uses an orifice without sheath flow. Turbulence
effects and non-uniform electric fields close to the orifice

sides and faces are probably responsible for the inaccuracies in

volume found when sheath flow is not used.

Figure & shows a blood sample from a patient with unusually small
red cells, who had received massive transfusions of normal cells.

The double peak Is clearly evident.

This type of instrument should have many applications In a

clinical laboratory.
Number of Cells

Figure 3

 

 

Volume

Volume Distribution of Red Cell sample measured in different ways.

ce
Number of Cells

 
 
 

Figure 4

Volume distribution of an abnormal Red Cell sample.

£@
-2h-

C. Use of flourescent techniques to test immunological

compatibility.

Work has continued on an automated version of the fluorochromatic
histocompatibility test. The pickup unit has worked
satisfactorily in conjunction with the optical and photoelectric
section of our IBM rapid cell spectrophotometer. Presently we are
completing a separate optical and photoelectric assembly so that
the instrument can work as a self-contained unit. This has been

supported by NIH Contract #GM17367.
-25-.

X. Optical Data Processing

One of the objectives of the NASA program for the exploration of
Mars is to obtain detailed images of thé planet from cameras
mounted on fly-by, orbiter, and lander spacecraft. Imagery data,
upon being telemetered back to Earth, is fed into a high-speed
computer for data manipulation and picture generation. Some of
the restoration operations required to yield pictures of high
quality require large amounts of computer time. As the realtime
demands of future missions can impose a heavy burden on available
computer capacity, we have performed experiments to investigate
the utility of analogue optical data processing procedures’ to

supplement the picture processing capability of the computer,

The equipment “assembled for this work is illustrated
schematically in Figure 5. Highly directed monochromatic light
from a 1 milliwatt continuous wave helium-neon laser is directed
through a shutter S to a microscope objective lens Lis A pinhole
aperture P, located at the focal point of Lie serves to exclude
stray radiation from the primary beam which passes through the
hole to a collimating lens L,- The light proceeds through two.
succeeding lens L, and Lie assumed here each to have focal length
f and to be separated from one another by a distance 2f. It
follows directly that if a phototransparency is inserted at 0, a
distance f in front of L,. then an image of the transparency wil]

be formed at |, a distance f behind L, The radiation pattern in
   

I9FT. STEEL H-BEAM

CUSHIONS _

BENCH

Fig.5 OPTICAL DATA PROCESSING SYSTEM

         
   
 

YZ

-97Z-
(a)

Input Transparency placed at 0.

Fig. 6 A portion of an image of

Mars

obtained in the

 

(b)

Output recorded at I.

course of the 1964 Fly-by Mission.

-lé-
~28-

the plane F located half way between L, and Ly is of an
especially interesting character for our purposes. There appears
in this plane a two dimensional Fourier transform of the
spatially variable transmissivity of the’ photo- transparency
placed at 0. The operational significance of this is that a
variety of useful transformations of the image appearing at ! can

be accomplished by placing masks of suitable configuration in the

plane F.

The application of the above system is then as follows: A
phototransparency representing a minimally processed computer
generated image of Mars is inserted at 0. A mask designed to
accomplish the desired transformation of the image is placed at
F, and a photo sensitive recording surface --Polaroid film in our
work-~- is placed at 1. The desired exposure is achieved with the

shutter.

Without elaborating on either the details of the radiation
pattern appearing at F or the selectton of an appropriate mask,
we illustrate two examples of operations that can be readily

performed.

Figure 6a shows an enlarged portion of an image of Mars obtained
from the 1964 Mariner 4& fly-by mission. the full image format
consists of an array of 200 by 200 distinct image points, each of

an intensity related to the amount of light recorded in the
(a)

Input Transparency placed at 0.

Fig. 7 An image of Mars recorded

 

(b)

Output recorded at I.

during the 1969 Mariner 6 Fly-by mission.

-6Z-
-30-

vicinity of the point. The overall effect is that of a half-tone
print such as is found in newspaper reproductions of photographs.
Normally the computer display operator would partially defocus
the image output tube to blend the individual dots to a somewhat
greater extent than shown here. In our demonstration we
accomplished a quite full blending by inserting an appropriate
mask at F. The output image recorded at I is illustrated in

Figure 6b.

Two features of Figure 6b bear comment. Firstly, the slightly
mottled character in the image results principally from the fact
that our mask was formed by cutting a sharply rounded hole in our
opaque sheet. This affect can be alleviated by utilization of a
"soft" edged mask. Secondly, imperfections in the input film are
exaggerated as indicated, for Instance, by the small black dots

beneath and about the dark area to the lower right of center.

Our second example, illustrated in Figure 7, involves an image
recorded during the 1969 Mariner 6 fly-by mission. The minimally
processed computer image shown in Figure 7a exhibits a form of
artifact common to the series, namely, harmonic "noise"
manifesting itself in this instance by some ten bands of
interference spread across the picture. The result of our effort
to suppress these bands by use of an appropriate F-ptane mask is

shown in Figure 7b.
-~31-

More sophisticated operations, of a pattern recognition type, can
be performed. We have not attempted these, nor do we believe it

likely that they can be developed in time to compete with digital

processing for MM 1971,
-32-

X!. Quasi=Microscope for Viking Mars Lander

The maximum spatial resolution capability of the facsimile camera
system proposed for the Mars Viking 1975 Lander is 1 mm. at a
minimum object range of 2 meter. Mission constraints for this
first Lander preclude the inclusion of a self-contained
microscope package. However, as there is great f[nterest In the
acquisition of microscopic data, we have given some thought’ to
the utilization of the proposed camera in a higher’ resolution

mode.

System reliability requirements preclude the addition of any
camera options that could in the event of malfunction of the
deployment system jeopardize the normal panoramic capability of

the system.

The simple observation that a magnifying eyeplece placed
approximately one focal length from an object but arbitrarily far
from the eye can provide substantial magnification, suggests a
possible approach. A remote auxilfary lens would enable the
camera to survey a smaller’ portion of its field in a

quasi-microscope mode.

We have studied both theoretically and experimentally the

features of such a system. We find for the proposed camera, ffor
-33-

example that a complementary F/1 auxillary lens of 40 mm diameter
placed at a range of 2 meter would enable acquisition of an
unvignetted image composed of 490 resolution elements at a
spatial resolution of 0.025 mm. - representing a 40-fold lateral

magnification relative to the unenhanced system.

General system equations have been derived and depth-of-field
relations have been established. We have examined an incidental
wide angle capability of the auxillary lens that offers the
option of quick scanning and/or monitoring over large angles and

areas.

It may be of more than academic interest to note a_ facsimile
microscope, patterned after the principle of the camera, can be
an extremely small device. An F/1] system consisting of a single
lens of arbitrarily small size and a pinhole focal plane aperture

can resolve down to one wavelength of light.
Xli. Mariner Mars 1971 Orbiter Photography

A system is being developed to manipulate digitized photographs
of Mars utilizing a time shared computer. The user is able to
interact with the system via a CRT display screen and a
typewriter keyboard. The system operates on a PDP=-10 computer
with 128,000 thirty-six bit words of core storage which is
augmented by a 20 million word disk system. The project is
directed toward increasing the speed and sensitivity with which
judgments can be made about features on the surface of Mars with

special emphasis on real time variation.

The user is able to select a region which is common to a pair of
images and request that the respective portions of the two images
be projected to a common point of view. Suitable geometric and
photometric transformations then generate two new images in which
common features are aligned to within the accuracy of the
spacecraft position and orientation data. Techniques also exist
for improving the alignment of the two transformed images’ to
within a fraction of a kilometer on the surface of Mars for’ the
1969 near-encounter photographs. After proper alignment, the. two
images can be analyzed to detect significant differences in the

two images of the same area. This technique is considered

fundamental to any significant variable features study.
The capability also exists to analyze the outlines of features

which are characterized by intensity differences in the’ image.

These features may be craters,. polar caps, sinuses or other
\

features of Mars. Work is also being done in recognizing,

classifying and measuring these features.
~36-
Xttt. REPORTS, PUBLICATIONS AND PAPERS
July 1, 1969 - Jury 1, 1970
PUBLICATIONS

1. G.W. Hodgson, E. Peterson, K.A. Kvenvolden, E. Bunnenberg,
B. Halpern, C. Ponnamperuma, "Search for Porphyrins in
Lunar Dust Returned from Apollo 11, Science 167, 763
(1970).

Ponnamperuma, K. Kvenvolden, S. Chang, R. Johnson,
Pollock, D. Philpott, I. Kaplan, J. Smith, J.W. Schopf,
Gehrke, G. Hodgson, |.A. Breger, B. Halpern, A. Duffield,
Krauskopf, E. Barghoorn, H. Holland, K. Keil, "Search

for Orzanic Compounds in the Lunar Dust from the Sea of
Tranquillity, Science 167, 760 (1970).

ces

3. B. Halpern, G.W. Yodgson, E. Bunnenbers, E. Peterson,
K. Kvenvolden, C. Ponnamperuma, "Search for Organic
Matter on the Moon," Geochemica et Cosmochimica Acta
Cin press, 1970).

4, WW. Pereira, V.A. Bacon, W. Patton and G.E. Pollock,
8. Halpern, "The Use of Phenylethylisocyanate in the
Optical Analysis of Asymmetric Secondary Alcohols by
G.L.C., Amal. Let 3, 23 (1970).

5. 1.S. Forrest, L.G. Brooks, S.D. Rose, B. Halpern, V.A,
Bacon, and J. Silberg, "Fluorescent Labelling of Psycho-
active Pruss, "Agressolozie (in press, 1970).

6. W. Pereira, V. Close, W. Patton, and B. Halpern, "Trans-
esterification with an Anion-Exchange Resin, Jour, Ore,
Chem. 34, 2032 (1969),

7. W. Pereira, V.A. Close, E£. Jellum, W. Patton and B.
Halpern, "Altcoholysis of the Merrifield-Type Pentide-
Polymer Bond with an Anion Exchange Resin," Aust, J.
Chem, 22, 1377-40 (1969).

8. 8B. Halpern, "Optical Activity" for "Exobfiology and the
Exploration of Mars," Applied Optics 8, 1349 (1969).

9. 8B. Halpern, W. Patton and P. Crabbe, "Cotton Effect of
Some Derivatives of Amino Acids and. Amino Alcohols,"
J. Chem, Soc, B, 1143 (1969).

10. E. Jellum, V.A. Close, W. Patton, W. Pereira, and 8.
Halpern, "A Gas-Liquid Chromatographic Method for the

Determination of Phenylalanine in Serum," Anal. Biochem,
31, 227 (1969).
11.

12.

13.

14,

15.

16,

17.

18.

19.

20.

21.

- 37 -

V. Bacon, E. Jellum, W. ‘Patton, W. Pereira, and B. Halpern,
"Peptide Sequencing by Low Resolution Mass Spectrometry I.
The Use of Acetylacetonyl Derivatives to !dentify N-
Terminal Residues," Biochem, and Biophys. Res. Comm. 37,
No. 6, 1969.

E., Jellum, V.A. Bacon, W. Patton, W. Pereira, Jr., and

B. Halpern, "Quantitative Determination of Biologically
Important Thiols and Disulfides by Gas-Liquid Chromato-
graphy," Anal. Biochem, 31, 339-347 (1969).

W. Patton, E. Jellum, D. Nitecki, W. Pereira and B.
Halpern, "The Modification of Phenylalanine Containing

‘Cyclo~Peptides with Nitrosy!l Chloride," Aust, J, Chem,

22, 2709 (1969).

D.P.L. Sachs, E. Jellum and 8. Halpern, "Netermination
of the Stereospecific Hydrolytic Action of Peptsin by
Nuclear Magnetic Resonance Spectroscopy," Biochim.

Biophys. Acta 198, 88 (1970).

S. Liebes, Jr., "Brightness - On the Ray Invariance of
B/n2," Am, Jour. Phys. 37(9), 932 (1969).

H.R. Hulett, W.A. Bonner, J. Barrett, L.A. Herzenberg,
"Cell Sorting: Automated Separation of Mammalian Cells
as a Function of Intracellular Fluorescence," Science
166, 747 (1969),

S. Liebes, Jr., "Gravitational Lens Simulator," Am, J.
Physics 37, 103 (1969).

H.R. Hulett, L.A. Herzenberg, W.A. Bonner, P.L. Wolf,
Rapid Cell Sorter - A New Tool for Cell Study with
Clinical Applications," Laboratory Investigations (in
press). To be presented at the 59th Annual Meeting of
the [International Academy of Pathology, Mar. 10-14, 1970,
St. Louis, Mo.

B. Halpern, W.E. Pereira, M.N. Solomon, E. Steed,
Submitted to Anal, Biochem. (1970).

B. Halpern, G.E. Pollock, "The Configuration of the
Olloisoleucine present in 'Maple Syrup urine disease’
Plasma", Submitted to Anal. Biochem, (1970).

J. Cymerman Craig, W.E. Pereira, BR. Halpern and J.W.
Westley. "Optical Rotatory Dispersion and Ahsolute
Confisuration XVII. G-Alkylphenyl acetic acids.
Submitted to Tetrahedron (1970).
22.

23.

26.

27.

28.

29.

30.

31.

32.

- 38 -

W. E. Pereira, M. Solomon, B. Halpern, "The Use of (+)
1-Pheny1]-2,2,2,-Trifluoroethyl-hydrazine in the Cptical
Analysis of Asymmetric Ketones by GLC". Submitted to
J. Ore, Chem. (1970).

WE. Pereira, B. Halpern, M.D. Solomon, A. M. Duffield,
"Electron Impact Promoted Fragmentation of Alkyl-N-
(1-Phenylethyl)-carbanates", Submitted to Ore. Mass,
Spectrom., (1970).

H.R. Yulett, "Non-enzymatic Hydrolysis of Adenosine
Phosphates". Nature 225,No0.5239:1248-1249 (1970).

W.E. Reynolds, V.A. Bacon, J.C. Bridges, T.C. Coburn,
B. Halpern, J. Lederberg, E£.C. Levinthal and £. Steed,
R.B. Tucker, "A Computer Operated Mass Spectrometer
System", Anal. Chem. (submitted, 1970).

W. E. Reynolds, "Instrumentation in a Time-Shared
Environment", Research/Development 21(4):20-26 (1970).

Lederberg, J., 1969. Mars through a crystal ball.
Applied Optics 8:1269-1270.

Schroll, Gustav, A. M. Duffield, Carl Djerassi, B.G.
Buchanan, G.L. Sutherland, E.A. Feigenbaum and

J. Lederberg, 1969. Applications of artificial intelligence
for chemical inference. II!. Aliphitic ethers diagnosed

by their low resolution mass spectra and NMR data.

J. Am. Chem. Soc, 91:7440.

Lederberg, J. Exobiology:search for evidence of life or
of biological habitats. Icarus (in press)

Lederberg, J., G.L. Sutherland, B.G. Buchanan and E.A.
Feigenbaum, 1969. A heuristic program for solving a
scientific inference problem: summary of motivation and
implementation. Stanford Artificial Intelligence Memo
AIM~104.,

A. Buchs, A.M. Duffield, G. Schroll, C. Djerassi, A.B.
Delfino, B.G. Buchanan, G.L. Sutherland, E.A. Feigenbaum,
and J. Lederberg, “Applications of Artificial Intelligence
for Chemical Inference IV. Saturated Amines Diagnosed by
Their Low Resolution Mass Spectra and Nuclear Magnetic
Resonance Spectra", 1970 (in press).

Y.M. Sheikh, A. Buchs, A.B. Delfino, G. Schroll, A.M.
Duffield, C. Djerassi, B.G. Buchanan, G.L. Sutherland,
E.A. Feigenbaum and J. Lederberg, "Applications of

Artificial Intelligence for Chemical Inference V. An
33.

34.

Approach to the Computer Generation of Cyclic Structures.
Differentiation Between All the Possible Isomeric Ketones
of Composition C6H100", Organic Mass Spectrometry, 1979
(in press).

A. Buchs, A.8. Delfino, A.M. Duffield, C. Djerassi,

B.G. Buchanan, E.A. Feigenbaum and J. Lederberg,
"Applications of Artificial Intelligence for Chemical
Inference VI. Approach to a General Method of Interpre-
ting Low Resolution Hass Spectra with a Computer",

Chem, Acta Helvetica, 1970 (in press).

G. Schroll, A.M. Duffield, Carl Djerassi, B.G. Buchanan,
G.L. Sutherland, E.A. Geigenbaum, and J. Lederberg,
"Applications of Artificial Intelligence for Chemical
Inference. 111. Aliphatic Ethers Diagnosed by Their
Low-Resolution Mass Spectra and Nuclear Magnetic Reso-
nance Data", J. Am, Chem. Soc, 91:7440 (1969),