PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Spectroscopy, Electron Energy-Loss”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 37 records · Page 2Linked to original sources

Presence of intranuclear inclusions in the principal cells of the epididymis of the garden dormouse Eliomys quercinus L.

Globular inclusions are present in the nuclei of the columnar epithelial cells lining the ductus epididymidis in the garden dormouse, Eliomys quercinus L. Such inclusions have been reported in the dog where they are very abundant, in the stallion and in man. They have been found to be absent from sheep, cat, rabbit, guinea pig, rat and mouse. We have now found them in the garden dormouse, an ascrotal hibernator, but have been unable to detect them in another hibernator, the thirteen-lined ground squirrel Citellus tridecemlineatus, this time a scrotal mammal. The globular inclusions are first seen in the perinuclear space formed by the two membranes of the nuclear envelope; they are then invested by the inner membrane of the nuclear envelope and transferred into the nucleus. They later disintegrate and the vacuole is finally left with an electron-transparent content. The number of inclusions varies with the hormonal status of the animal: the granules are always present as soon as there is a rise in plasma testosterone. Electron energy-loss spectroscopy in the electron microscope (EELS) shows that inclusions contain C, N, O, Fe, Mg, P and S. This suggests that the intranuclear granules are made of protein cross-linked by disulphur bonds and contain iron, magnesium and phosphorus.

Animals↗

Quantitative analysis of electron energy-loss spectra from ultrathin-sectioned biological material. I. Optimization of the background-fit with the use of Bio-standards.

A computer program for quantitative spectral analysis is proposed for the elemental analysis of biological material by electron energy-loss spectroscopy in a conventional transmission electron microscope, the Zeiss EM902. Bio-standards are used to test the performance of this program. The application of a simplex optimization method for curve-fitting is proposed to separate the ionization edge from the background. Making use of Ce-, Ca- and Fe-bio-standards, this method is compared with Egerton's well-known two-area method.

Calcium↗

Electronic core level microanalyses and microcopies in multipurpose apparatus.

Using an instrument equipped with two electron guns, an electron analyzer, and a Si(Li) diode detector, we developed microanalytical techniques based on inner-shell electron excitations by incident electrons and X-rays, that is, electron energy-loss spectroscopy (EELS) in the reflection mode; electron probe microanalysis (EPMA) and X-ray appearance potential spectroscopy (XAPS); electron-induced Auger electron spectroscopy (e-AES); X-ray photoelectron spectroscopy (XPS), X-ray absorption spectroscopy (XAS); X-ray induced AES (XAES), X-ray fluorescence analysis (XRF), and scanning X-ray radiography (SXR). The corresponding characteristic images (including X-ray microradiography and X-ray photoelectron microscopy) were obtained in the scanning mode. The principle of the apparatus is described. Each spectroscopy and microscopy is illustrated by an example. Their performance and limits are discussed.

Electron Probe Microanalysis↗

Analysis of the calcium distribution in predentine by EELS and of the early crystal formation in dentine by ESI and ESD.

Predentine is a collagen-rich extracellular matrix between the odontoblasts and the dentine with a width of about 15-20 microns. Electron energy-loss spectroscopy of rat incisors shows a significantly higher calcium content in the predentine at the predentine-dentine border than in the middle region of the predentine. At the predentine-dentine border in the dentine, the calcium and the phosphate groups combine to form apatite crystallites. Electron spectroscopic diffraction with zero-loss filtering revealed that the earliest crystallites contain only Debye-Scherrer rings of apatite, which are fewer in number and more diffuse than the diffraction rings from the mature crystallites. We therefore conclude that the early crystallites still contain lattice defects, which are annealed out to some degree with crystal growth. Electron spectroscopic imaging with zero-loss filtering also showed that the earliest crystallites are chains of dots (or small islands); they build up strands composed of islands, which rapidly acquire a needle-like character and coalesce laterally to form ribbon-or plate-like crystallites. The parallel strands sometimes appear to reinforce the macroperiod of the collagen microfibrils (67 nm) by tiny holes without any crystal-substance lined up perpendicular to the parallel strands of the crystallites.

Animals↗

Improved preservation of phospholipid-rich multilamellar bodies in conventionally embedded mammalian lung tissue--an electron spectroscopic study.

Different conventional methods of tissue processing were studied to determine the extent to which phospholipid-rich multilamellar bodies of pulmonary alveolar epithelial type II cells of the pig were preserved. Prolonged treatment with half-saturated aqueous uranyl acetate yielded excellent results on the stabilization of the multilamellar substructure, irrespective of whether glutaraldehyde-paraformaldehyde or glutaraldehyde-tannic acid was used as a primary fixative. The lamellar periodicities were observed to be 5.5-6.1 nm. Differences in the phosphorus distribution among several types of lipid bodies of alveolar epithelial type II cells were studied by means of electron spectroscopic imaging and electron energy-loss spectroscopy. Multilamellar bodies gave phosphorus signals which were significantly higher than those obtained from granular regions of composite bodies, whereas homogeneous bodies gave phosphorus signals which were even lower than those obtained from mitochondria, endoplasmic reticulum membranes or ribosomes.

Animals↗

Electron microscopy, electron diffraction, and element analysis of wet biological specimens.

Temperature controlled differentially pumped environmental chambers now allow more routine examination of wet specimens in the electron microscope. A sensitive test of their efficiency is the ability to provide high resolution electron diffraction patterns from wet, unfixed protein microcrystals. Fortunately, wet specimens can be prepared with only a few tens of nanometers thickness of remaining water, so extraneous electron scattering by liquid water can be kept to a minimum. It still remains to be determined whether microprobe analysis (X-ray or electron energy-loss spectroscopy) using wet specimens gives better element localization in cells than the current freezing methods. More extensive comparisons are also required of the ultrastructural preservation and visibility of macromolecules immersed in a thin layer of water vs immersion in a thin layer of amorphous ice. However, the recent introduction of commercial forms of the necessary equipment now make these comparisons more feasible.

Freeze Drying↗

Three-dimensional structure of frozen-hydrated paracrystals of myosin rod.

Myosin rod, the tail fragment of myosin, aggregates into various structures at physiological ionic strength. One form, the type II paracrystal, a thin, ribbon-like species, has recently been studied using conventional electron microscopy and was shown to possess two-dimensional order and, further, is likely to be useful in the investigation of the arrangement of myosin molecules in the backbone of the vertebrate muscle thick filament (Ward & Bennett, 1989). We have now examined this aggregate in the frozen-hydrated state by cryo-electron microscopy. Image analysis indicated that the projected structure has the same p12, plane group symmetry as seen after negative staining. A three-dimensional map, calculated from projections, indicated that the structure comprises a bilayer arrangement of strands of grouped rod molecules, with the strands parallel in each layer. The layers themselves are related by screw symmetry. Strands in adjacent layers have opposing polarity with their long axes at an angle of 32 degree to each other. Protein density measurements, carried out on unstained specimens using electron energy-loss spectroscopy, showed that the 44 X 13 X 13 nm unit cell is composed of 40% protein. The density measurements indicated that 9-12 rod molecules pass through each strand. Modelling rod molecules with 43 nm parallel overlaps in a body-centered tetragonal lattice produced a strand that compared favorably with the reconstructed strand. The size and content of these strands suggests that they are analogous to subfilaments observed in the vertebrate myosin thick filament.

Animals↗

EELS data acquisition, processing and display for the Zeiss CEM 902 based on LOTUS 1-2-3: application examples from a biological system and inorganic transition metal compounds.

A personal computer combined with LOTUS 1-2-3 software, including the RS232 module of LOTUS MEASURE and a 12-bit ADC, has been used for data acquisition of electron energy-loss spectroscopy (EELS) spectra with the Zeiss CEM 902. The internal macro language of LOTUS 1-2-3 allows a menu-driven procedure. Macro-programs partly combined with external FORTRAN programs can be chosen from the menu for background subtraction, removal of multiple scattering effects by deconvolution, elemental quantification and several utilities. For special applications or conditions the macro programs can easily be modified. Spectra from crystals of two inorganic transition metal compounds, ruthenium trichloride and vanadium disulphide, and from a biological sample are presented as examples of the application of this software.

Animals↗

Ultrastructural and electron spectroscopic analyses of cyanobacteria and bacteria.

The extracellular sheath material and some intracellular cell components of cyanobacteria and phosphate-accumulating sewage bacteria were analysed by electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS). The specimens were embedded in water-soluble Nanoplast resin without any previous fixation and ultrathin sections were examined in a Zeiss CEM 902 microscope. A high sulphur content was detected in the inner sheath of the cyanobacterium Gloeothece. The elemental composition of some cell components and inclusion bodies, such as carboxysomes and cyanophycin, was determined by ESI and EELS. In addition, the phosphate content in specific granules of phosphate-accumulating sewage bacteria was estimated by EELS and nuclear magnetic resonance spectroscopy.

Bacteria↗

Calcium transport mechanism in molting crayfish revealed by microanalysis.

Crayfish provide a good model in which to study the transport mechanism of Ca ions. During the molting stage, decalcified Ca ions are transferred into the blood and accumulate in the gastrolith epithelium, after which a gastrolith is formed on the surface of the epithelium. The gastrolith is dissolved in the stomach after molting, and the Ca is reabsorbed and redistributed throughout the newly formed exoskeleton. We studied the mechanism of Ca transport by cytochemical precipitation of Ca ions and by electron microanalysis, including X-ray microanalysis (EDX) and electron energy-loss spectroscopy (EELS), with a computer. In EDX analysis, the fine precipitates of K-antimonate in the gastrolith mitochondria clearly defined Ca with antimony; we also observed a large amount of Ca-oxalate in the mitochondria, and Ca-K X-ray pulses were clearly defined. Ca-K X-rays were also detected from fresh freeze-substituted mitochondria. Finally, we succeeded in taking a Ca-L EELS image from the mitochondria of fresh freeze-substituted thin sections. Only a very small amount of Ca was detected from the cell membrane and other organelles. Ca-adenosine triphosphatase (ATPase) and Mg-ATPase activity was also very clearly demonstrated in the mitochondria. These enzymes may play an important role in Ca metabolism.

Adenosine Triphosphatases↗

The use of parallel EEL spectral imaging and elemental mapping in the rapid assessment of anti-cancer drug localization.

Both electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS) have great potential for use in several areas of cancer research. In biologically targeted radiotherapy, cytotoxic drug therapy and boron neutron capture therapy the effectiveness of many drugs is often critically dependent upon the intracellular localization of the agent employed. We describe the use of parallel EEL spectral imaging to assess the penetration and location of the iodine-containing drug meta-iodobenzyl guanidine, of potential value in targeted radiotherapy, and for the rapid detection of boron within borate-adsorbed polystyrene beads, of potential value in boron neutron capture therapy. We also describe elemental mapping of boron following low-temperature embedding. These results show how the techniques could be applied to many forms of cancer research by discussing the validity and limitations of the techniques experimentally. We also provide an outline of other areas in this field which could benefit from the future application of ESI and EELS.

3-Iodobenzylguanidine↗

Quantitative analysis of electron energy-loss spectra from ultrathin-sectioned biological material. II. The application of bio-standards for quantitative analysis.

Electron energy-loss spectroscopy (EELS) has been used to determine elemental concentrations in biological specimens, consisting of ultrathin-sectioned cells and tissues. Chelex100-based Ca- and Fe Bio-standards are used for elemental quantification to establish iron and calcium concentrations. These Bio-standards, as well as the biological materials, are treated in a standard EM procedure such that 'known' and 'unknown' sites are located in one cross-section. Uncertainties and variabilities present in the equations for calculating the concentration in the 'unknown' site (determined by comparing simplex-fitted EEL spectra from Bio-standards with those from tissue) are outlined in two examples. Using an H+ Bio-standard, the matrix composition of such biological cell material is analysed, leading to values which approach each other closely. Quantitative EELS, using Chelex100-based Bio-standards, is advocated.

Animals↗

Reflection high resolution analytical electron microscopy: a technique for studying crystal surfaces.

Reflection electron microscopy (REM), reflection high energy electron diffraction (RHEED), reflection electron energy-loss spectroscopy (REELS), and energy dispersion x-ray spectroscopy (EDX) have been comprehensively used as a technique, termed reflection high resolution analytical electron microscopy (RHRAEM), for studying the structures of the bulk crystal GaAs (110) surfaces by transmission electron microscopy (TEM). The simultaneous observations of surface topography imaging, the surface diffraction mechanism with RHEED, surface atomic inner-shell excitations with REELS, and surface chemical compositions with EDX provide a systematic description of the atomic structure and chemical structure of the surface. The surface channelling effect has been observed in GaAs (110) with REELS, which may provide a basis for localizing surface foreign atoms with ALCHEMI. The theoretically predicted surface-resonance wave has been observed directly in the RHEED pattern; the surface-captured Bragg reflection wave have been identified. It is shown that surface chemical compositions can be determined by analyzing the EDX spectra obtained in the REM case. Finally, the surface monolayer resonance characteristic of the RHRAEM has been confirmed by calculations with dynamical RHEED theory.

Arsenic↗

Energy-loss near-edge fine structure and compositional profiles of cryomilled oxide-dispersion-strengthened aluminum.

Spatially resolved electron energy-loss spectroscopy (SREELS) makes possible the identification of small second-phase particles in an oxide-dispersion-strengthened aluminum alloy produced by mechanical alloying in a liquid-nitrogen slurry. Near-edge fine structure profiles based on aluminum L23 features that are characteristic of Al-Al or Al-O bonding yield the metal/oxide fraction as a function of probe position. Profiles of nitrogen fraction are derived from linear sequences of spectra so that detection limits can be analyzed at each point in a scan. Quantitative analysis of these linear profiles shows that particles with diameters in the range 10 to 30 nm are rich in nitrogen. A combination of statistical analysis of background removal results with visual inspection of derivative spectra is used to estimate detection limits.

Aluminum↗

Analysis of directly frozen macromolecules and tissues in the field-emission STEM.

A VG Microscopes HB501 field-emission high-resolution scanning transmission electron microscope (STEM) was used to image and analyse rapidly frozen, isolated macromolecules and small organelles in tissue cryosections. Dark-field images were obtained from frozen-hydrated microtubules demonstrating that sufficient contrast is available to reveal structural information. The samples were subsequently freeze-dried in the STEM and low-dose (approximately 10(3) e/nm2) dark-field mass maps were recorded with single electron sensitivity. Elemental analysis of individual macromolecules was achievable at high dose using parallel-detection electron energy-loss spectroscopy, albeit with some structural degradation. Detection of copper (320 atoms) in di-decameric haemocyanin molecules was easily within the limits of sensitivity. Elemental analysis of hydrated cryosections is limited by radiation damage to a resolution of approximately 1 micron2. For freeze-dried sections, however, the high probe current and stable cold stage of the HB501 STEM allow energy-dispersive X-ray (EDX) microanalysis of low elemental concentrations in highly localized subcellular volumes. EDX spectra from cryosections of cerebellar cortex show that a 100-s analysis time is sufficient to quantify the calcium content of 400-nm2 regions within Purkinje cell dendrites with an uncertainty of +/- 2 mmol/kg dry weight, equivalent to +/- 12 atoms.

Animals↗

Contrast in the electron spectroscopic imaging mode of a TEM. IV. Thick specimens imaged by the most-probable energy loss.

When the zero-loss transmission falls below 10(-3) for biological sections of mass-thickness greater than 70 micrograms/cm2, the energy window in the electron spectroscopic imaging (ESI) mode of an energy-filtering electron microscope (EFEM) can be shifted to the most-probable energy loss of the electron energy-loss spectrum. This enables mass-thicknesses up to 150 micrograms/cm2 or thicknesses of 1.5 microns to be examined. Electron energy-loss spectra of thick carbon films calculated by a Fourier method agree with experimental spectra. Measurements of the electron energy-loss spectroscopy and ESI image intensities with an additional platinum film confirm a scattering model for the calculation of the image intensity. This model considers the angular broadening at the most-probable energy loss by introducing an effective illumination aperture of the order of the full-width at half-maximum of the angular distribution.

Fourier Analysis↗

Electron spectroscopic study (ESI, EELS) of Nanoplast-embedded mammalian lung.

The potential of Nanoplast melamine resin embedding for the study of mammalian lung parenchyma was examined by means of electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS). Samples were either fixed with glutaraldehyde-paraformaldehyde or glutaraldehyde-tannic acid, or were directly transferred to the embedding medium without prior fixation. Organic dehydrants, as well as fixatives containing heavy metals and stains, were omitted. A very high level of ultrastructural detail of chromatin, ribosomes, mitochondria and plasma membranes was achieved by ESI from the Nanoplast-embedded samples. The most prominent gain in ultrastructural detail was achieved when moving from an energy loss just below the L2,3 edge of phosphorus at 132 eV to an energy loss just beyond this edge. This reflects the prominent P L2,3 edge observed by EELS of Nanoplast-embedded samples in comparison with conventionally processed samples. Thus, taking into account possible sectioning artefacts, excellent heterochromatin images which rely on the phosphorus distribution can be obtained from Nanoplast-embedded samples by computer-assisted analysis of electron spectroscopic images. In this respect glutaraldehyde-paraformaldehyde fixation is preferable to glutaraldehyde-tannic acid fixation because the presence of silicon, revealed by EELS, in tannic-acid-fixed samples may introduce artefacts in phosphorus distribution images obtained by the three-window method because of the close proximity of the L2,3 edges of silicon and phosphorus.

Animals↗

The influence of objective lens aberrations in energy-loss spectrometry.

The conditions under which the energy resolution and collection efficiency in electron energy-loss spectrometry are limited by the spherical and chromatic aberrations of the objective lens have been studied. It is shown that, for the optimum settings of the pre-spectrometer optics, the energy resolution will be of the same order in diffraction mode as it is in magnification mode. The influence of the aberrations on the practice of energy-loss spectroscopy is discussed, and it is demonstrated that the chromatic aberration can act as a broadband filter for energy-loss electrons.

Electron Probe Microanalysis↗