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Results for “phase-contrast microscopy”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
NASA NTRS1974

Studies on Auditory and Vestibular End Organs and Brain Stem Nuclei

Cats were exposed to tones of 125, 1000, 2000, and 4000 Hz at sound pressure levels in the range 120 to 157.5 db, and for durations of one hour (1000, 2000, 4000 Hz) or four hours (125 Hz). Pure tone audiograms were obtained for each animal before and after exposure. Cochleas of animals were examined by phase-contrast microscopy. Extent of inner ear damage and range of frequencies for which hearing loss occurred increased as exposure tone was decreased in frequency. For example, exposure to 4000 Hz produced damage in a restricted region of the cochlea and hearing loss for a relatively narrow range of frequencies; exposure to 125 Hz produced wide-spread inner ear damage and hearing loss throughout the frequency range 125 to 6000 Hz.

Ades, H. W.↗
NASA NTRS1992

Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans

A rapid, sensitive, inexpensive in situ hybridization technique, using 30-mer 16S rRNA probes, can specifically differentiate two closely related Bacillus spp., B. polymyxa and B. macerans. The 16S rRNA probes were labeled with a rhodamine derivative (Texas Red), and quantitative fluorescence measurements were made on individual bacterial cells. The microscopic fields analyzed were selected by phase-contrast microscopy, and the fluorescence imaging analyses were performed on 16 to 67 individual cells. The labeled 16S rRNA probe, POL, whose sequence was a 100% match with B. polymyxa 16S rRNA but only a 60% match with B. macerans 16S rRNA, gave quantitative fluorescence ratio measurements that were 34.8-fold higher for B. polymyxa cells than for B. macerans cells. Conversely, the labeled probe, MAC, which matched B. polymyxa 16S rRNA in 86.6% of its positions and B. macerans 16S rRNA in 100% of its positions, gave quantitative fluorescence measurements that were 59.3-fold higher in B. macerans cells than in B. polymyxa cells. Control probes, whose 16S rRNA sequence segment (P-M) was present in both B. polymyxa and B. macerans as well as a panprokaryotic probe (16S), having a 100% match with all known bacteria, hybridized equally well with both organisms. These latter hybridizations generated very high fluorescence signals, but their comparative fluorescence ratios (the differences between two organisms) were low. The control paneukaryotic probe (28S), which had less than 30% identity for both B. macerans and B. polymyxa, did not hybridize with either organism.

Non-NASA Center↗
NASA NTRS1992

Colour thresholding and objective quantification in bioimaging

Computer imaging is rapidly becoming an indispensable tool for the quantification of variables in research and medicine. Whilst its use in medicine has largely been limited to qualitative observations, imaging in applied basic sciences, medical research and biotechnology demands objective quantification of the variables in question. In black and white densitometry (0-256 levels of intensity) the separation of subtle differences between closely related hues from stains is sometimes very difficult. True-colour and real-time video microscopy analysis offer choices not previously available with monochrome systems. In this paper we demonstrate the usefulness of colour thresholding, which has so far proven indispensable for proper objective quantification of the products of histochemical reactions and/or subtle differences in tissue and cells. In addition, we provide interested, but untrained readers with basic information that may assist decisions regarding the most suitable set-up for a project under consideration. Data from projects in progress at Tulane are shown to illustrate the advantage of colour thresholding over monochrome densitometry and for objective quantification of subtle colour differences between experimental and control samples.

NASA Discipline Neuroscience↗
NASA NTRS2016

Biomolecular Analysis Capability for Cellular and Omics Research on the International Space Station

International Space Station (ISS) assembly complete ushered a new era focused on utilization of this state-of-the-art orbiting laboratory to advance science and technology research in a wide array of disciplines, with benefits to Earth and space exploration. ISS enabling capability for research in cellular and molecular biology includes equipment for in situ, on-orbit analysis of biomolecules. Applications of this growing capability range from biomedicine and biotechnology to the emerging field of Omics. For example, Biomolecule Sequencer is a space-based miniature DNA sequencer that provides nucleotide sequence data for entire samples, which may be used for purposes such as microorganism identification and astrobiology. It complements the use of WetLab-2 SmartCycler"TradeMark", which extracts RNA and provides real-time quantitative gene expression data analysis from biospecimens sampled or cultured onboard the ISS, for downlink to ground investigators, with applications ranging from clinical tissue evaluation to multigenerational assessment of organismal alterations. And the Genes in Space-1 investigation, aimed at examining epigenetic changes, employs polymerase chain reaction to detect immune system alterations. In addition, an increasing assortment of tools to visualize the subcellular distribution of tagged macromolecules is becoming available onboard the ISS. For instance, the NASA LMM (Light Microscopy Module) is a flexible light microscopy imaging facility that enables imaging of physical and biological microscopic phenomena in microgravity. Another light microscopy system modified for use in space to image life sciences payloads is initially used by the Heart Cells investigation ("Effects of Microgravity on Stem Cell-Derived Cardiomyocytes for Human Cardiovascular Disease Modeling and Drug Discovery"). Also, the JAXA Microscope system can perform remotely controllable light, phase-contrast, and fluorescent observations. And upcoming confocal microscopy capability will allow for optical sectioning of biological tissues to determine microanatomical localization of biomarkers. Furthermore, NASA's geneLAB effort addresses integration of genomic, epigenomic, transcriptomic, proteomic and metabolomic datasets, by applying an innovative open source science platform for multi-investigator high throughput utilization of the ISS. In sum, the expanding ISS capability for analysis of biomolecules is enabling innovative research in a broad spectrum of areas such as cellular and molecular biology, biotechnology, tissue engineering, biomedicine, and Omics, providing manifold benefits for humanity.

Guinart-Ramirez, Y.↗
NASA NTRS1989

Ecology and life history of an amoebomastigote, Paratetramitus jugosus, from a microbial mat: new evidence for multiple fission

Five microbial habitats (gypsum crust, gypsum photosynthetic community, Microcoleus mat, Thiocapsa scum, and black mud) were sampled for the presence of the euryhaline, rapidly growing amoebomastigote, Paratetramitus jugosus. Field investigations of microbial mats from Baja California Norte, Mexico, and Salina Bido near Matanzas, Cuba, reveal that P. jugosus is most frequently found in the Thiocapsa layer of microbial mats. Various stages of the life history were studied using phase-contrast, differential-interference, and transmission electron microscopy. Mastigote stages were induced and studied by electron microscopy; mastigotes that actively feed on bacteria bear two or more undulipodia. A three-dimensional drawing of the kinetid ("basal apparatus") based on electron micrographs is presented. Although promitoses were occasionally observed, it is unlikely that they can account for the rapid growth of P. jugosus populations on culture media. Dense, refractile, spherical, and irregular-shaped bodies were seen at all times in all cultures along with small mononucleate (approximately 2-7 micrometers diameter) amoebae. Cytochemical studies employing two different fluorescent stains for DNA (DAPI, mithramycin) verified the presence of DNA in these small bodies. Chromatin-like material seen in electron micrographs within the cytoplasm and blebbing off nuclei were interpreted to the chromatin bodies. Our interpretation, consistent with the data but not proven, is that propagation by multiple fission of released chromatin bodies that become small amoebae may occur in Paratetramitus jugosus. These observations are consistent with descriptions of amoeba propagules in the early literature (Hogue, 1914).

Non-NASA Center↗
NASA NTRS2002

Tetracycline rapidly reaches all the constituent cells of uropathogenic Escherichia coli biofilms

We have developed a method for visualizing Escherichia coli cells that are exposed to tetracycline in a biofilm, based on a previous report that liposomes containing the E. coli TetR(B) protein fluoresce when exposed to this antibiotic. By our method, cells devoid of TetR(B) also exhibited tetracycline-dependent fluorescence. At 50 microg of tetracycline ml(-1), planktonic cells of a uropathogenic E. coli (UPEC) strain developed maximal fluorescence after 7.5 to 10 min of exposure. A similar behavior was exhibited by cells in a 24- or 48-h UPEC biofilm, as examined by confocal laser microscopy, regardless of whether they lined empty spaces or occupied densely packed regions. Further, a comparison of phase-contrast and fluorescent images of corresponding biofilm zones showed that all the cells fluoresced. Thus, all the biofilm cells were exposed to tetracycline and there were no pockets within the biofilm where the antibiotic failed to reach. It also appeared unlikely that niches of reduced exposure to the antibiotic existed within the biofilms.

Non-NASA Center↗
NASA NTRS1999

Microscopy of Analogs for Martian Dust and Soil

The upcoming Mars 2001 lander will carry an atomic force microscope (AFM) as part of the Mars Environmental Compatibility Assessment (MECA) payload. By operating in a tapping mode, the AFM is capable of sub-nanometer resolution in three dimensions and can distinguish between substances of different compositions by employing phase-contrast imaging. Phase imaging is an extension of tapping-mode AFM that provides nanometer-scale information about surface composition not revealed in the topography. Phase imaging maps the phase of the cantilever oscillation during the tapping mode scan, hence detecting variations in composition, adhesion, friction, and viscoelasticity. Because phase imaging highlights edges and is not affected by large-scale height differences, it provides for clearer observation of fine features, such as grain edges, which can be obscured by rough topography. To prepare for the Mars 01 mission, we are testing the AFM on a lunar soil and terrestrial basaltic glasses to determine the AFMOs ability to define particle shapes and sizes and grain-surface textures. The test materials include the Apollo 17 soil 79221, which is a mixture of agglutinates, impact and volcanic beads, and mare and highland rock and mineral fragments. The majority of the lunar soil particles are less than 100 microns in size, comparable to the sizes estimated for Martian dust. The terrestrial samples are millimeter size basaltic glasses collected on Black Pointe at Mono Lake, just north of the Long Valley caldera in California. The basaltic glass formed by a phreatomagmatic eruption 13,000 years ago beneath a glacier that covered the Mono Lake region. Because basaltic glass formed by reworking of pyroclastic deposits may represent a likely source for Martian dunes, these basaltic glass samples represent plausible analogs to the types of particles that may be studied in sand dunes by the 01 lander and rover. We have used the AFM to examine several different soil particles at various resolutions. The instrument has demonstrated the ability to identify parallel ridges characteristic of twinning on a 150-micron plagioclase feldspar particle. Extremely small (10-100 nanometer) adhering particles are visible on the surface of the feldspar grain, and appear elongate with smooth surfaces. Phase contrast imaging of the nanometer particles shows several compositions to be present. When the AFM was applied to a 100-micron glass spherule, it was possible to define an extremely smooth surface.E Also visible on the surface of the glass spherule were chains of 100-nanometer- and-smaller impact melt droplets. Additional information is contained in the original extended abstract.

Anderson, M. A.↗
NASA NTRS2005

Phase-Imaging with a Sharpened Multi-Walled Carbon Nanotube AFM Tip: Investigation of Low-k Dielectric Polymer Hybrids

Phase shift tapping mode scanning force microscopy (TMSFM) has evolved into a very powerful technique for the nanoscale surface characterization of compositional variations in heterogeneous samples. Phase shift signal measures the difference between the phase angle of the excitation signal and the phase angle of the cantilever response. The signal correlates to the tip-sample inelastic interactions, identifying the different chemical and/or physical property of surfaces. In general, the resolution and quality of scanning probe microscopic images are highly dependent on the size of the scanning probe tip. In improving AFM tip technology, we recently developed a technique for sharpening the tip of a multi-walled carbon nanotube (CNT) AFM tip, reducing the radius of curvature of the CNT tip to less than 5 nm while still maintaining the inherent stability of multi-walled CNT tips. Herein we report the use of sharpened (CNT) AFM tips for phase-imaging of polymer hybrids, a precursor for generating nanoporous low-k dielectrics for on-chip interconnect applications. Using sharpened CNT tips, we obtained phase-contrast images having domains less than 10 nm. In contrast, conventional Si tips and unsharpened CNT tips (radius greater than 15 nm) were not able to resolve the nanoscale domains in the polymer hybrid films. C1early, the size of the CNT tip contributes significantly to the resolution of phase-contrast imaging. In addition, a study on the nonlinear tapping dynamics of the multi-walled CNT tip indicates that the multi-walled CNT tip is immune to conventional imaging instabilities related to the coexistence of attractive and repulsive tapping regimes. This factor may also contribute to the phase-contrast image quality of multi-walled CNT AFM tips. This presentation will also offer data in support of the stability of the CNT tip for phase shift TMSFM.

Nguyen, Cattien V.↗