scholarly journals UV transmission microscopy – optical imaging without glass

2021 ◽  
pp. 4-15
Author(s):  
Jonathan Crowther ◽  
Author(s):  
William Krakow

For the best transmission electron microscopes now available, the phase contrast mechanism must be used in the interpretation of the optical imaging process producing high resolution micrographs. In the following paragraphs, some techniques are discussed which minimize image degradation and yield improvements in resolution and/or contrast so that the information content in the image can be clearly displayed.


Author(s):  
E. P. Abrahamson II ◽  
M. W. Dumais

In a transmission microscopy study of iron and dilute iron base alloys, it was determined that it is possible to preserve specimens for extended periods of time. Our specimens were prepunched from 5 to 8 mil sheet to microscope size and annealed for several hours at 700°C. They were then thinned in a glacial acetic-12 percent perchloric acid solution using 10 volts and 20 milliamperes, at a temperature of 8 to 14°C.It was noted that by the use of a cold stage, the same specimen can be observed for periods up to one week without excess contamination. When removal of the specimen from the column becomes necessary, it was observed that a specimen may be kept for later observation in 1,2 dichloroethene or methanol for periods in excess of two weeks.


Author(s):  
W. R. Duff ◽  
L. E. Thomas ◽  
R. M. Fisher ◽  
S. V. Radcliffe

Successful retrieval of the television camera and other components from the Surveyor III spacecraft by the Apollo 12 astronauts has provided a unique opportunity to study the effects of a known and relatively extensive exposure to the lunar environment. Microstructural effects including those produced by micro-meteorite impact, radiation damage (by both the solar wind and cosmic rays) and solar heating might be expected in the materials used to fabricate the spacecraft. Samples received were in the form of 1 cm2 of painted unpainted aluminum alloy sheet from the top of the camera visor (JPL Code 933) and the sides (935,936) and bottom (934) of the lower camera shroud. They were prepared for transmission electron microscopy by first hand-grinding with abrasive paper to a thickness of 0.006". The edges were lacquered and the sample electropolished in 10% perchloric methanol using the “window” method, to a thickness of ~0.001". Final thinning was accomplished by polishing 3 mm punched disks in an acetic-phosphoric-nitric acid solution.


Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.


Author(s):  
M. H. Wheeler ◽  
W. J. Tolmsoff ◽  
A. A. Bell

(+)-Scytalone [3,4-dihydro-3,6,8-trihydroxy-l-(2Hj-naphthalenone] and 1,8-di- hydroxynaphthalene (DHN) have been proposed as intermediates of melanin synthesis in the fungi Verticillium dahliae (1, 2, 3, 4) and Thielaviopsis basicola (4, 5). Scytalone is enzymatically dehydrated by V. dahliae to 1,3,8-trihydroxynaphthalene which is then reduced to (-)-vermelone [(-)-3,4- dihydro-3,8-dihydroxy-1(2H)-naphthalenone]. Vermelone is subsequently dehydrated to DHN which is enzymatically polymerized to melanin.Melanin formation in Curvularia sp., Alternaria sp., and Drechslera soro- kiniana was examined by light and electron-transmission microscopy. Wild-type isolates of each fungus were compared with albino mutants before and after treatment with 1 mM scytalone or 0.1 mM DHN in 50 mM potassium phosphate buffer, pH 7.0. Both chemicals were converted to dark pigments in the walls of hyphae and conidia of the albino mutants. The darkened cells were similar in appearance to corresponding cells of the wild types under the light microscope.


Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).


Author(s):  
R. E. Heffelfinger ◽  
C. W. Melton ◽  
D. L. Kiefer ◽  
W. M. Henry ◽  
R. J. Thompson

A methodology has been developed and demonstrated which is capable of determining total amounts of asbestos fibers and fibrils in air ranging from as low as fractional nanograms per cubic meter (ng/m3) of air to several micrograms/m3. The method involves the collection of samples on an absolute filter and provides an unequivocal identification and quantification of the total asbestos contents including fibrils in the collected samples.The developed method depends on the trituration under controlled conditions to reduce the fibers to fibrils, separation of the asbestos fibrils from other collected air particulates (beneficiation), and the use of transmission microscopy for identification and quantification. Its validity has been tested by comparative analyses by neutron activation techniques. It can supply the data needed to set emissions criteria and to serve as a basis for assessing the potential hazard for asbestos pollution to the populace.


Author(s):  
H. Koike ◽  
T. Matsuo ◽  
K. Ueno ◽  
M. Suzuki

Since the identification of single atoms was achieved by Crewe et al, scanning transmission microscopy has been put into pratical use. Recently they applied this method to the quantitative mass analysis of DNA.As pointed out previously the chromatic aberration which decreases the image contrast and quality, does not affect a scanning transmission image as it does a conventional transmission electron microscope image. Thus, the STEM method is advantageous for thick specimen. Further this method employs a high sensitive photomultiplier tube which also functions as an image intensifier. This detection method is effective for the observation of living specimens or easily damaged specimens. In this respect the scanning transmission microscope with high accelerating voltage is necessary.Since Uyeda's experiments of crystalline materials, many workers have been discussed how thick specimens can be observed by CTEM. With biological specimens, R. Szirmae reported on the decrease in the image contrast of rabbit psoas muscle sections at various accelerating voltages and specimen thicknesses.


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