AUTORADIOGRAPHY OF FATIGUE FRACTURE IN ALUMINUM
Autoradiography of fatigue fracture in aluminum
RADIOGRAPHY↗
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High-Resolution Autoradiography
This investigation was made to adapt wet-process autoradiography to metallurgical samples to obtain high resolution of segregated radioactive elements in microstructures. Results are confined to development of the technique, which was perfected to a resolution of less than 10 microns. The radioactive samples included carbon-14 carburized iron and steel, nickel-63 electroplated samples, a powder product containing nickel-63, and tungsten-185 in N-155 alloy.
Towe, George C↗
Photographic image intensification by autoradiography
Images on developed photographic emulsions can be significantly intensified by making the image silver radioactive and exposing a second emulsion to this radiation. Because the print, an autoradiograph, reproduces the original with significant increases in density and contrast, the method can be used to enhance underexposed emulsions and to increase the limits of photographic detection. This paper reports a new, practical chemical method for activating the silver, using thiourea labeled with sulfur-35, a process convenient for use in photographic laboratories. Speed increases of 13 and contrast increases of 18 were achieved with Kodak Plus-X film in this initial phase.
Askins, B. S.↗
Post-development image intensification by autoradiography
A method of post-development intensification of photographic images using a newly developed autoradiographic procedure is described. Radioactive Thiourea-S35 is combined with silver in the developed emulsion, which is then contact printed with a receiver emulsion. The beta decay from S35 produces an enhanced autoradiographic image. The laboratory technique is described and results of astronomical applications are presented.
Askins, B. S.↗
High-resolution autoradiography
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Towe, George C↗
Gene activity during germination of spores of the fern, Onoclea sensibilis: RNA and protein synthesis and the role of stored mRNA
Pattern of 3H-uridine incorporation into RNA of spores of Onoclea sensibilis imbibed in complete darkness (non-germinating conditions) and induced to germinate in red light was followed by oligo-dT cellulose chromatography, gel electrophoresis coupled with fluorography and autoradiography. In dark-imbibed spores, RNA synthesis was initiated about 24 h after sowing, with most of the label accumulating in the high mol. wt. poly(A) -RNA fraction. There was no incorporation of the label into poly(A) +RNA until 48 h after sowing. In contrast, photo-induced spores began to synthesize all fractions of RNA within 12 h after sowing and by 24 h, incorporation of 3H-uridine into RNA of irradiated spores was nearly 70-fold higher than that into dark-imbibed spores. Protein synthesis, as monitored by 3H-arginine incorporation into the acid-insoluble fraction and by autoradiography, was initiated in spores within 1-2 h after sowing under both conditions. Autoradiographic experiments also showed that onset of protein synthesis in the cytoplasm of the germinating spore is independent of the transport of newly synthesized nuclear RNA. One-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis of 35S-methionine-labelled proteins revealed a good correspondence between proteins synthesized in a cell-free translation system directed by poly(A) +RNA of dormant spores and those synthesized in vivo by dark-imbibed and photo-induced spores. These results indicate that stored mRNAs of O. sensibilis spores are functionally competent and provide templates for the synthesis of proteins during dark-imbibition and germination.
NASA Discipline Number 29-20↗
Quantitative analysis of autoradiographic image intensification using Thiourea-S35
Photographic images enhanced by the method of Thiourea-S35 autoradiography are evaluated in terms of signal-to-noise ratio, detective quantum efficiency (DQE), and Wiener spectrum analysis using digitized images. It is determined that the original signal-to-noise ratio is not degraded by the intensification process which allows an increase in the practical working DQE as a function of density. These results apply at all spatial frequencies that were tested. The advantage given by autoradiography is the ability to produce usable images from emulsions originally exposed to the low densities corresponding to maximum DQE and movement of faint image densities above the level of the threshold for detection.
Askins, B. S.↗
In situ trace element microanalysis
The use of particle-track-radiography and X-ray- fluorescence techniques in the in situ measurement of trace (less than 1000 ppm) elements in single mineral phases of polished sections is surveyed, and examples of their application to ordinary, carbonaceous and enstatite chondrites are provided. Radiographic methods surveyed include fission-track radiography (for U, Th, and Pu-244), alpha radiography using nuclear reactions (for Li and B), alpha autoradiography (for Bi and Pb), and beta autoradiography (for several elements in synthetic or biological samples). Two X-ray-fluorescence methods are compared: (1) photon-induced X-ray emission (PIXE), and (2) the potential use of synchrotron radiation. The latter is shown to allow much greater sensitivity than current PIXE technology and a much broader range of elements than particle-track radiography: the ppm analysis of 10-micron grains for all elements heavier than Na. These advantages are seen as balancing the high cost of accelerator use.
Burnett, D. S.↗
Cell Kinetic and Histomorphometric Analysis of Microgravitational Osteopenia: PARE.03B
Previous methods of identifying cells undergoing DNA synthesis (S-phase) utilized H-3 thymidine (3HT) autoradiography. 5-Bromo-2'-deoxyuridine (BrdU) immunohistochemistry is a nonradioactive alternative method. This experiment compared the two methods using the nuclear volume model for osteoblast histogenesis in two different embedding media. Twenty Sprague-Dawley rats were used, with half receiving 3HT (1 micro Ci/g) and the other half BrdU (50 microgram/g). Condyies were embedded (one side in paraffin, the other in plastic) and S-phase nuclei were identified using either autoradiography or immunohistochemistry. The fractional distribution of preosteoblast cell types and the percentage of labeled cells (within each cell fraction and label index) were calculated and expressed as mean q standard error. Chi-Square analysis showed only a minor difference in the fractional distribution of cell types. However, there were significant differences (p less than 0.05) by ANOVA, in the nuclear labeling of specific cell types. With the exception of the less-differentiated A+A'cells, more BrdU label was consistently detected in paraffin than in plastic-embedded sections. In general, more nuclei were labeled with 3H-thymidine than with BrdU in both types of embedding media. Labeling index data (labeled cells/total cells sampled x 100) indicated that BrdU in paraffin, but not plastic gave the same results as 3HT in either embedding method. Thus, we conclude that the two labeling methods do not yield the same results for the nuclear volume model and that embedding media is an important factor whenusing BrdU. As a result of this work, 3HT was chosen for used in the PARE.03 flight experiments.
Roberts, W. Eugene↗
Cell Kinetic and Histomorphometric Analysis of Microgravitational Osteopenia: PARE.03B
Previous methods of identifying cells undergoing DNA synthesis (S-phase) utilized 3H-thymidine (3HT) autoradiography. 5-Bromo-2'-deoxyuridine (BrdU) immunohistochemistry is a nonradioactive alternative method. This experiment compared the two methods using the nuclear volume model for osteoblast histogenesis in two different embedding media. Twenty Sprague-Dawley rats were used, with half receiving 3HT (1 micro-Ci/g) and the other half BrdU (50 micro-g/g). Condyles were embedded (one side in paraffin, the other in plastic) and S-phase nuclei were identified using either autoradiography or immunohistochemistry. The fractional distribution of preosteoblast cell types and the percentage of labeled cells (within each cell fraction and label index) were calculated and expressed as mean +/- standard error. Chi-Square analysis showed only a minor difference in the fractional distribution of cell types. However, there were,significant differences (p less than 0.05) by ANOVA, in the nuclear labeling of specific cell types. With the exception of the less-differentiated A+A' cells, more BrdU label was consistently detected in paraffin than in plastic-embedded sections. In general, more nuclei were labeled with 3H-thymidine than with BrdU in both types of embedding media (Fig 2.). Labeling index data (labeled cells/total cells sampled x 100) indicated that BrdU in paraffin, but not plastic gave the same results as 3HT in either embedding method. Thus, we conclude that the two labeling methods do not yield the same results.
Roberts, W. Eugene↗
Distinguishing Indigenous from Contaminating Microorganisms in Rock Samples from a Deep Au Mine in South Africa
The concentration and distribution of microbial biomass within deep subsurface rock strata is not well known To date, most analyses are from water samples and a few cores. Hand samples, block samples and cores from an actively mined Carbon Leader ore zone at 3.2 kilometers depth were collected for microbial analyses. The Carbon Leader was comprised of quartz, S-bearing aromatic hydrocarbons, Fe(III) oxyhydroxides, sulfides, uraninite, Au and minor amounts of sulfate. The porosity of the ore was 1% and the maximum pore throat diameter was less than 0.1 microns; whereas, the porosity of the adjacent quartzite was .02 to .9% with a maximum pore throat diameter of 0.9 microns. Rhodamine dye, fluorescent microspheres, microbial enrichments, autoradiography, phospholipid fatty acid (PLEA) and 16S rDNA analyses were performed on these rock samples and the mining water. The date indicate that the levels of solute contamination less than 0.01% for pared rock samples. Despite this low level of contamination, PLEA, microbial enrichment, DNA and tracer analyses and calculations indicate that most of the viable microorganisms in the Carbon Leader represent gram negative aerobic heterotrophs and ammonia oxidizers that are phylogenetically identical or closely related to service water microorganisms. These microbial contaminants probably infiltrated the low permeability rock through mining-induced microfractures. Geochemical data also detected drilling water in a fault zone approx. 1 meter behind the rock face encountered during coring. The mining induced macrofractures that are common at these great depths act as pathways for the drilling water borne microorganisms into the lower temperature zone that extends several meters into rock strata from the rock face. Combined PLEA and T- RFLP analyses of the service water and Carbon Leader samples indicate that the concentration of indigenous microorganisms was less than 10(exp 2) cells/gram. Such a low concentrations result from the submicron pore throat diameters. PLFA. SO4-35 autoradiography and tracer analyses indicate that the bounding quartzite contains thermophilic sulfate reducing bacteria at 10(exp 3) cells/gram that are not attributable to drilling water contamination. The microorganisms may be surviving on sulfate generated by oxidation of sulfide by radiolytic reactions resulting from the high U concentration in the ore zone. The presence of up to 8,000 ppm of Fe(III) oxyhydroxides in the host rock will also act to recycle sulfide generated by the sulfate reducing bacteria into sulfate. The activity of these sulfate-reducing bacteria may be enhanced by mining induced fracturing which can propagate up to 40 meters into virgin rock where the temperatures are ca. 50 C, and decrepitate of sulfate rich fluid inclusions. In ultra deep mines, judicious application of tracers and multiple microbial characterization techniques can distinguish microbial contamination caused by the near field fracturing and drilling water migration from the indigenous microbial communities in rock strata. The importance of far field fracturing on indigenous microbial communities, however, remains unknown.
Onstott, T. C.↗
Approaches in the determination of plant nutrient uptake and distribution in space flight conditions
The effective growth and development of vascular plants rely on the adequate availability of water and nutrients. Inefficiency in either the initial absorption, transportation, or distribution of these elements are factors which impinge on plant structure and metabolic integrity. The potential effect of space flight and microgravity conditions on the efficiency of these processes is unclear. Limitations in the available quantity of space-grown plant material and the sensitivity of routine analytical techniques have made an evaluation of these processes impractical. However, the recent introduction of new plant cultivating methodologies supporting the application of radionuclide elements and subsequent autoradiography techniques provides a highly sensitive investigative approach amenable to space flight studies. Experiments involving the use of gel based 'nutrient packs' and the radionuclides calcium-45 and iron-59 were conducted on the Shuttle mission STS-94. Uptake rates of the radionuclides between ground and flight plant material appeared comparable.
manned↗
Nucleic acid and protein synthesis during lateral root initiation in Marsilea quadrifolia (Marsileaceae)
The pattern of DNA, RNA, and protein synthesis during lateral root initiation in Marsilea quadrifolia L. was monitored by autoradiography of incorporated of 3H-thymidine, 3H-uridine, and 3H-leucine, respectively. DNA synthesis was associated with the enlargement of the lateral root initial prior to its division. Consistent with histological studies, derivatives of the lateral root initial as well as the cells of the adjacent inner cortex and pericycle of the parent root also continued to synthesize DNA. RNA and protein synthetic activities were found to be higher in the lateral root initials than in the endodermal initials of the same longitudinal layer. The data suggest a role for nucleic acid and protein synthesis during cytodifferentiation of a potential endodermal cell into a lateral root initial.
Non-NASA Center↗
Novel use of positively charged nylon transfer membranes for trapping indoleacetic acid or other small anions during efflux from plant tissues
Positively charged nylon blotting membranes were used as an anion binding medium to trap [14C]indoleactic acid (IAA) as it exited cells at the basal ends of Coleus blumei L. stem and Zea mays L. coleoptile segments. Autoradiography was used to visualize where the [14C] that moved out of the cut ends was localized on the nylon membrane. Diffusion of [14C]IAA from the initial point of contact with the nylon membrane was minimal. Comparison of the autoradiograms with anatomical tissue prints of the cut ends of the segments was used to determine what tissues participate in IAA movement. The results of these initial studies were consistent with other reports suggesting that [14C]IAA movement was primarily associated with vascular tissues in both C. blumei stems and corn coleoptiles, but the resolution was not sufficient to identify which vascular tissues were involved in IAA transport. With further refinements, this technique could also be used for studying the movement of other small charged molecules through plant tissues.
NASA Discipline Plant Biology↗
Embryonic lung morphogenesis in organ culture: experimental evidence for a proteoglycan function in the extracellular matrix
The lung rudiment, isolated from mid-gestation (11 day) mouse embryos, can undergo morphogenesis in organ culture. Observation of living rudiments, in culture, reveals both growth and ongoing bronchiolar branching activity. To detect proteoglycan (PG) biosynthesis, and deposition in the extracellular matrix, rudiments were metabolically labeled with radioactive sulfate, then fixed, embedded, sectioned and processed for autoradiography. The sulfated glycosaminoglycan (GAG) types, composing the carbohydrate component of the proteoglycans, were evaluated by selective GAG degradative approaches that showed chondroitin sulfate PG principally associated with the interstitial matrix, and heparan sulfate PG principally associated with the basement membrane. Experiments using the proteoglycan biosynthesis disrupter, beta-xyloside, suggest that when chondroitin sulfate PG deposition into the ECM is perturbed, branching morphogenesis is compromised.
NASA Discipline Number 93-10↗
The Influence of Surface Coatings on Exo-Electron Emission During Fatigue
A survey of exo-electron emission literature has shown that the mechanism of exo-emission phenomena has yet to be ascertained. Also, there is probably no unique mechanism that will apply to all the exo-emission observations reported. For exo-emission to occur, trapping centers must first be formed to store electrons. Once such centers are formed, they can be made to give up their trapped electrons by the stimulation of light or oxygen. Future investigations will employ light-transparent films for collecting exo-electrons. Also, a carefully controlled experiment is necessary to detect the emission of exo-electrons from the surface of fatiguing metals. Investigations will include high-vacuum fatigue tests, autoradiography, and solid state collector experiments, and will provide a basis for future research on semiconductor layer collection.
COATING↗
Autoradiographic localization of radioactivity in rat brain after injection of tritiated sex hormones.
Sex hormone uptake localization in brains of rats determined with high spatial resolution by autoradiography, using tritiated testosterone and estradiol
Pfaff, D. W.↗
Analysis of T4 phage proteins. I
T4 phage proteins radioisotopical examination, determining quantitative analysis and molecular weight by autoradiography
Cone, R.↗