Document rpyq6RzREbevvYqv1gpjd3D7J
FILE NAME: CTFA (CTFA) DATE: 1974 April DOC#: CTFA025 DOCUMENT DESCRIPTION: Article from Trade Journal-American Laboratory
i -*
I5T
Contents
DEPARTMENTS COMING COVER
Asbestos, 13
BY WALTER C. MC CRONE AND IAN M. STEWART
Optical information processing in the characterization of microscope images, 23
BY B. H. KAYE AND A. G. NAYLOR
Nutrition for chemists, 29
BY ROGER J . WILLIAMS
Humans, the mammalian mutants, 32
BY IRWIN STONE
A high intensity transmitted light base for dark-field stereomicroscopy, 40
BY GENE E. SCHLUETER
Aspiration and dispensing system for hand-held pipets, 47
BY JAMES F. HOLSINGER
Desk-top scanning electron microscopes, 51
BY DONALD J . EVINS AND ROBERT J . ENGLE
Physics of the cell membrane: part one, the role of double-bond energy states, 59
BY A. KEITH BREWER AND RICHARD A. PASSWATER
Consultation microscopy, a review, 77
BY BERNARD FRIEDMAN
Analytical scanning electron microscopy, 83
BY B. W. GRIFFITHS
Toward automating radioimmunoassay, 91
BY D. J . MARSHALL
The research marketplace, 1974-1980,103
BY WILLIAM N. WHAM AND KENNETH S. HALABY
Editor's page, 8 Computer interfacing: the view from the transmission line, 45
BY DAVID G. LARSEN AND PETER R. RONY
Laboratory profile: Walter C. McCrone Associates, Inc., 97 Scientific economics, 109 New products, 111 Available literature, 129 Advertising index, 136 Author's format, RSC
The Delaware State College drug analysis program Differential light scattering analyses Determination of organic vapors in industrial atmospheres Gas-liquid chromatography of nucleic acid constituents High speed gel permeation chromatography Vapor phase pyrolysis and gas chromatographic analysis of fire retardant materials containing polyvinyl chloride Design concepts of a processor-based gas chromatograph
Photomicrograph of vitamin C as seen through a polarized microscope. Courtesy of Sidney Braginsky.
VOLUME 6, NUMBER 4
Published monthly by International Scientific Communications, Inc., Greens Farms, Connecticut 06436, U.S.A. Phone 203 255-2645. Controlled circulation postage paid at Dallas, Texas. Subscriptions available: $18.00 per year, add $3.60 for postage to all foreign countries; $1.75 for single copies, add $0.40 extra to all foreign countries. International Scientific Communications, Inc. assumes no responsibility for the state ments and opinions advanced by the contributors.
Copyright 1974 by International Scientific Communications, Inc. All rights reserved.
Reproduction in whole or in part without written permission from International Scientific
Communications, Inc. is prohibited.
albybl 6 (4) 1-138 (1974)
4 : APRIL 1974
American Laboratory
Asbestos
By W alter C. McCrone and Ian M. Stewart
In m a y 1968, Dr. Irving J. Selikoff of the Mount Sinai Hos pital in New York established his position as the Rachel Carson
followed up by the FDA, Depart ment of Labor, and other inter ested parties, most responsible companies are now doing their
of the industrial hygiene world. A best to minimize, if not eliminate,
series of
NewYorker articleassbaetstos as a health hazard.
that time was based largely on Dr.
The microscopist has an im
SelikofFs experiences in the study portant role in this program to
of asbestosis, lung and stomach control asbestos as a contaminant
cancer, mesothelioma, and other in the air we breathe, the fluids
causes of death related to ex we drink, and the food we eat. He
posure to asbestos. His careful must be able to identify asbestos
analyses of mortality of industrial fibers and quantify their size and
workers in known asbestos fiber number. There are problems be
contaminated areas are convinc cause there are other fibrous sub
ing arguments that asbestos is stances confusingly similar to as
one of the most deadly industrial bestos, and there are several dif
hygiene hazards.
ferent types of asbestos ( Table 1),
The reputation of large indus differing in optical properties and
trial concerns has not always been composition and possibly in the
good when faced with evidence degree of their hazard to humans.
that their plant environments may
Analytical methods must be
be contributing to worker disabil sensitive enough for the identifica
ities, if not death. However, large tion of single fibers. Of course,
ly through Dr. Selikoff's effort. only a microscope and a trained
Dr. McCrone is Scientific Adviser and Mr. Stewart is Manager, Electron Optics Group, McCrone Associates,
Inc.
microscopist can do this. There are methods, e.g., x-ray diffrac tion and DTA. that identify min ute percentages of asbestos in
Chrysotile fiber bundle.
Table 1 Composition
Serpentine
CHRYSOTILE 3 M g 0 2 S i0 2*
2H20
ACTINOLITE 2C aO *4M gO F e 0*8S i02H20
Asbestos
Amphiboles
TREM OLITE
2CaO-5MgO 8S i02*H 20
ANTHOPHYLLITE 7Mg0` 8Si02H20
Specific gravity Crystal system Refractive indices
2.36-2.5 monoclinic 1.49-1.57
3.03-3.5 monoclinic 1.62-1.68
2.9-3.2 monocHnic 1.60-1.65
2.85-3.5 orthorhombic
1.60-1.66
AMOSITE
5-5FeO1*5MgO 8S i02H20 2.6-3.0 monoclinic 1.66-1.70
CROCIDOLITE
Na20Fe20 3 3Fe0*8Si02
H20 3.0-3.45 monoclinic 1.69-1.71
AMERICAN LABORATORY : 13
ASBESTOS continued
Table 2 Asbestos type
Chrysotlle Anthophyllite Amosite Crocidolite
Dispersion staining colors for asbestos
Refractive index liquid
Dispersion staining color
Parallel
Perpendicular
1.560 1.610 1.670 1.700
light blue blue-green red magenta magenta
magenta golden yellow golden yellow blue magenta
small samples, but only the mi-
croscopist can hope to identify in
dividual fibers. Fortunately, the
crystallographic properties of the
different types of asbestos are suf
ficiently distinct to enable certain
identification. If the fibers are
thicker than about 0.5
light
microscopy suffices; smaller fibers
require the electron microscope.
Identification of the larger fi bers by light microscopy is based on refractive index measurement using a polarizing microscope. The easiest way to apply refrac tive index measurement to asbes tos fiber identification is by use of dispersion staining, an optical staining procedure. This is based on annular or central screening of
the objective aperture in common with specific refractive index me dia, such as the Cargille liquids. Under these conditions each type of asbestos shows two character istic dispersion staining colors, one for the light vibration direc tion parallel, and another perpen dicular, to the fiber length (but only in one particular refractive
index liquid). Table 2 shows the colors obtained with several kinds of asbestos when mounted in ap propriate Cargille refractive index liquids. Different colors or no colors are observed if other than the specified liquids are used for each particular asbestos sample. Finding small percentages of very fine asbestos fibers requires care ful application of the method. In particular, intense Kohler illumi nation with carefully centered substage and objective apertures is essential. A careful microscopist should, however, be able to detect 0.01% of >0.5 firnthick fibers by this technique. Light microscopy is ideal for rapid identification of asbestos in most powdered samples: dust,
mortar, insulation or wallboard,
mineral samples such as talc, etc.
When the presence of asbestos in
a liquid medium, such as a water
supply, soft drink, beer, or pa
renteral solution is suspected, one
must assume most of the fibers,
if not all, will be too fine to ob
serve by light microscopy. One
turns, then, to the transmission
electron microscope (
7).
The transmission electron micro
scope (TEM) is superior to the
scanning electron microscope for
two reasons. Smaller fibers can
be observed and identified by
TEM, and selected area electron
diffraction (SAED) is more de
pendable for identification of as
bestos than the elemental analysis
capability of the SEM.
Figure 1 Model 200KV transmission electron microscope.
EXTENDING MAN'S VISION IN THE SEAS
The seas are our new frontier. From an underw ater Nikonos
Scientists are exploring the camera to the compact Model vast resources of th e oceans, H microscope, we are producnot only for links to our origins, instrum ents used by our but for new sources of food. scientist/conservationists to
Many scientists believe th at ^ ^ c o n t i n u a t i o n of the
our beleagured planet cannot forever support the popula tion explosion.
The w arning voices of these men are at last being heard. Mankind is slowly awakening to the frightening realization that even our seas cannot w ithstand the absorption of man's w aste without dire consequences
species' Producing the finest instru ments available for science and industry is a constant challenge.
W herever essential research is being conducted; on land, beneath the seas, or in space-- we are meeting that challenge.
Extending your vision is our main concern.
Nikon is in the forefront of undersea research technology. The finest optics in the \\ orlo.
Nikon Instrument Division, Ehrenreich Photo-Optical Industries,
(N ikon)
--p
US
623 Stewart Ave., Garden City, N.Y.
Tel. (516) 248-5200.
In Canada, Anglophoto Ltd., Ontario.
Nikon SMZ Stereo Zoom Microscope Total magnification range. 4X to 120X. Provides 5time zoom range without change in focus. Excep tional wide range of accessories. Reader Card fc 107
Nikim Epi-Illuminsitor Fits any standard Nikon microscope. Designed for observation of opaque specimens. Provides colli mated on-axis illumination with field and aperture control. Dark field reflected light systems available. Reader Card *08
Nikon Ki Compact Versatile laboratory microscope with built-in transformer, continuously variable lieht intensity control, and inbase volt meter. Features brilliant Koehler type illuminauon and full range of Nikon accessories. Reader C ards 109
AMERICAN LABORATORY : 15
BAUSCH & LOMB
BaId Ian MICROSCOPE One of a kind
I Optical A rchitecture ... so outstanding in Bausch & Lomb's new Balplan Mi
croscope, makes using this research quality instrument a
delightful experience. Interchanging modules, attaching
accessories, so quick and uncomplicated; focusing, so
fine--so smooth; your most vital investigations become
pleasanter, less fatiguing, most rewarding.
Balplan's modular system of integrated components is
mounted on a stand that features an exclusive, suspended
inner arm. Pile as many accessories on the stand as you
need, the optics, as part of the vibration-free inner arm,
will still maintain drift-free, accurate focus.
Focusing is controlled by concentric, comfortably low-
positioned knobs. Fine focus is graduated in single mi
crons, allowing you to dissect visually each layer of the
specimen with ease. A complete revolution of the focus
ing knob moves the stage 100 microns.
With the transformer housed in the base and with front
push-button controls all circuitry is enclosed.
The line cord plugs into the base at the rear
while accessory cords plug into the upper back
area of the stand. Hazardous, bothersome dan
gling wires are eliminated.
Notably improving performance, the patented
slide holder on the mechanical stage is spring
mounted downwards to hold the slide in con stant contact with the stage surface. Nosepieces for 2, 3, 4 or 5 objectives are reversible,
interchangeable, parcentered and accurately position ob jectives with a ball detent.
Accessories for visual and photographic use are all de
signed to be compatible within the Balplan system. To top it off--There's a choice of interchangeable
heads, including a dual-viewing adapter with illuminated pointer.
That, in brief, is the architecture of
, the one-
w.
of-a-kind new concept in a competitively priced micro
LOOK INTO BALPLAN. No other microscope system offers
so much for so Httie. . . a premium quality microscope at up
to half the price of another brand of comparable quality. See
for yourself in a live demonstration in your lab with vour own
specimens.
3
scope with models for clinical, research and educational applications.
Send for new 24-page full color catalog 31-2411 and our free demonstration offer.
TM. Blplan >5 a Bausch & Lomb Tradename
BAUSCH & LOMB. SCIENTIFIC OPTICAL PRODUCTS DIVISION. 38804 BAUSCH STREET, ROCHESTER N Y 14602 Circle Reader Service Card No. 13 for information; No. 14 for dem onstration.
ASBESTOS continued
A sample of water of known
volume is filtered through the fin
est membrane filter. A section of
filter approximately 2-3 mm2 is
then placed particle side down on
a previously carbon-coated elec
tron microscope grid, and the
membrane filter is dissolved using
acetone in a Soxhlet extractor.
The sample grids are examined in
our laboratory with a JEM 200
electron microscope, using an ac
celerating voltage of 150 kv at a
magnification of 17,000X on the
M3 range of the instrument. This
magnification is chosen to focus
the intermediate lens aperture in
line with the specimen plane; thus,
a SAED pattern of the fiber is ob
tained with no other adjustments
to the microscope. Tn this way it
is possible to spot check the dif
fraction pattern of individual fi
bers rapidly (
Figure2 and
The total number of asbestos
fibers is related to a definite area
of the filter, and hence a definite
volume of sample, by counting fi
bers over a known number of
TEM grid squares (having known
areas). The length and width of
each asbestos fiber is recorded.
Interpolation from the screen
scribed at half-centimeter intervals
allows an accuracy of measure
ment on the screen of approxi
mately 0.05 cm, or about 3 4
nm. Measurements of the indi
vidual fibers are read into a tape
recorder; this permits the operator
to devote full attention to the
screen, precludes duplicate count
ing of single fibers, prevents miss
ing fibers, and maintains the op
erator's dark accommodation.
The recorded data are trans
cribed onto punch-tape and com
puter processed to give listings of
the length, width, and aspect ratio
of the fibers, together with a com
puted mass of each fiber calcu
lated on the basis of density D
and dimensions L and
X
assigns the longest dimension to the fiber length and excludes all fibers with an aspect ratio below three.
This procedure, designed by Ian Stewart of our laboratory, has been applied with excellent suc cess to a number of water samples containing large numbers of amphiboles per liter but very low weight percentages. Most of these samples show no asbestos fibers by light microscopy. It is there fore important to specify the method of analysis when stating analytical results. The same sam ple of talc could be analyzed by different methods with very dif ferent results, e.g.:
1. X-ray diffraction shows no chrysotile or amphiboles.
L X
W2).Density values for th2e. A phase-contrast method
3d).ifferent asbestos types are given (similar to the NIOSH procedure
in Table 1. Also presented in the to determine the number of fibers
computer print-out is the num per milliliter of air) shows 1500
ber of fibers per liter of water, fibers greater than 5/*m/m3 of
the size distribution of the fibers sample.
based on length and width, and the distribution of fibers by aspect ratio. The program automatically
3. The scanning electron mi croscope with energy dispersive x-ray analyzer shows about 104
AMERICAN LABORATORY : 17
UniS ude
Manual & Motor Driven
Linear Slides
for Instruments & Machines
Sssk-
RAPIO ADVANCE
TYPICAL
56 Manual models,
ASBESTOS continued
fibers/liter suspected of being am-
phibole asbestos.
4. Dispersion staining shows
150 anthophyllite fibers/gram of
sample.
5. Transmission electron mi
croscopy shows 5 X 10 antho
phyllite fibers/liter of sample.
The AEI Scientific Apparatus,
Ltd. EMMA-4 (
4) only
recently installed in our labora
tory will undoubtedly be the ideal
instrument for the detection and
identification of very fine asbestos
fibers. A combination transmis
sion electron microscope and
electron microprobe analyzer
(EM A), it enables the micro-
scopist to see the smallest fiber
and to identify it by diffraction
pattern and elemental analysis.
Although a given sample mounted
on a TEM grid can be examined
successively by TEM and EMA,
the advantage of being able to examine many particles in a sam ple with assurance of one-to-one correspondence of particle size, shape, diffraction pattern, and elemental analysis is obvious. We have already applied this instru ment to some asbestos samples with excellent results, and expect it will be the instrument of choice for such samples in our labora tory.
The monitoring of asbestos as a contaminant in our environment is an excellent example of one important problem solved by microscopy. The importance of choosing the proper analytical tool and having a well-trained op erator is also emphasized by this application to asbestos detection and identification.
Figure 4 EMMA-4, a combined transmission electron microscope and electron microprobe analyzer.
y L M E X INC
P.O. BOX 38 E. BLOOMFIELD, NY 14443 Telephone 3 1 5 /657 -61 51
Circle Reader Service Card No. 69
18 : APRIL 1974