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At least 1,153 records · Page 64Linked to original sources
PubMed1997

In vitro calcified tendon collagen: an atomic force and scanning electron microscopy investigation.

Atomic force microscopy (AFM), scanning electron microscopy and X-ray energy dispersive spectroscopy have been performed on decalcified turkey tendons submitted to in vitro calcification in order to investigate the morphology and the surface relationships between the inorganic phase and the collagen fibres during deposition and compare with those found for physiologically calcified samples. 'Tapping mode' AFM was used to reduce the vertical force applied to the samples, which were examined without any preparation. A further characterization has been carried out by means of X-ray diffraction, infrared absorption and chemical analyses. The observations indicate that the inorganic phase deposited on collagen fibres during in vitro calcification is poorly crystalline B carbonated apatite. The composition, structure and dimensions of apatitic crystallites, as well as their orientation with respect to collagen fibrils, are very similar to those characteristic of physiologically calcified tissues. However, the crystallites seem to be nucleated on the fibril surface, without appreciably affecting the molecular packing of collagen.

Animals↗
PubMed1979

Structural states of myelin observed by x-ray diffraction and freeze-fracture electron microscopy.

Coordinated freeze-fracture electron microscopy and x-ray diffraction were used to visualize the morphological relation between compacted and native period membrane arrays in myelinated nerves treated with dimethylsulfoxide (DMSO). Comparison of x-ray diffraction at room temperature and at low temperature was used as a critical measure of the extent of structural preservation. Our x-ray diffraction patterns show that in the presence of cryoprotective agents, it is possible to preserve with only small changes the myelin structure which exists at room temperature. These changes include a slight increase in packing disorder of the membrane, a small, negative thermal expansion of the membrane unit, and some reorganization in the cytoplasmic half of the bilayer. The freeze-fracture electron microscopy clearly demonstrates continuity of compact and native period phases in DMSO-treated myelin. Finally, the use of freezing to trap the transient, intermediate structure during a structural transition in glycerol is demonstrated.

Animals↗
PubMed1996

[Electron microscopy for the studies of membrane transport proteins].

Electron microscopy gives the real images of individual protein molecules, providing unique means in the structural research of membrane-transport proteins. Various techniques for the preparation of specimens for transmission electron microscopy were briefly described. Emphases were put on the recent progress of quick-freeze cryo-electron microscopy, including cryotransfer and freeze-fracture deep-etch replication. Exploiting the advantage of the latter technique, we visualized the in situ structure of inositol 1,4,5-trisphosphate receptor molecules in the smooth-surfaced endoplasmic reticulum (sER) membrane of the Purkinje neuron in bovine cerebellum. The tetrameric receptor molecules were unexpectedly small and formed a two-dimensional crystalline array on the surface of the cisternal stacks derived from sER. Its structural characteristics were discussed in comparison with the ryanodine-receptor, the other intracellular Ca2+ -channel.

Animals↗
PubMed1977

Scanning and transmission electron microscopy of the nasal mucosa.

We attempted to study the nasal mucosa of children (both normal and diseased) under light microscopy followed by both scanning and transmission electron microscopy. Combined methods of microscopy are valuable tools in establishing the diagnosis and lead to a better understanding of disease and its management. For this reason we found it necessary to study the ultrastructural details of the cells of the nasal mucosa. Through scanning electron microscopy we studied the functional aspects and the nature of the surface of these cells. Ultrastructural details of these cells were studied with the aid of transmission electron microscopy.

Adolescent↗
PubMed1993

Scanning electron microscopy of synaptonemal complexes.

The novel application of scanning electron microscopy to study whole-mount surface-spread synaptonemal complex complements of rye (Secale cereale) and rat (Rattus norvegicus) is described. Scanning electron microscopy is able to resolve the third dimension in such preparations and improve the tracing of the continuity of lateral elements without losing information that could be obtained by conventional transmission electron microscopy. This improvement is likely to benefit detailed studies of chromosome synapsis and karyology, and may provide a means of circumventing technical obstacles inhibiting the use of surface-spreads as substrates for in situ hybridization under the electron microscope.

Animals↗
PubMed1985

Small round cell neoplasms: can electron microscopy and immunohistochemical studies accurately classify them?

The answer to the question posed in the title, "Small Round Cell Neoplasms: Can Electron Microscopy and Immunohistochemical Studies Accurately Classify Them?", is obviously "yes"; but a qualified yes--generally yes, perhaps with expertise usually yes, but never just plain yes. Some cases certainly will defy the best attempts even of the most expert in the application of these "special" techniques. And embarrassing as it may be for those of us infatuated with the latest technology to admit, it is with the difficult case especially that old-fashioned technology so often must be depended upon. In his excellent recent appraisal of the role of a variety of special techniques in this application, Triche offers the following comment: "Overall, electron microscopy is probably the most universally useful of all diagnostic techniques other than light microscopy in round cell tumors." The data from our studies certainly point to the same conclusion. With each of the tumors, electron microscopy demonstrated itself to be more reliable than immunohistochemistry. Electron microscopy offers not only greater sensitivity and specificity, but also greater versatility. Immunohistochemistry allows hypothesis testing only. Electron microscopy, on the other hand, can provide answers even when the right questions are not being asked. For example, if a particular small round cell tumor under investigation happens in actuality to represent something other than the neuroblastoma which it is being considered (e.g., a granulocytic sarcoma, liposarcoma, Wilm's tumor, etc.), electron microscopy can reveal this fact, but a neuron-specific enolase stain cannot. Parenthetically, it should also be said that electron microscopy has proven particularly well suited to the examination of fine-needle aspiration specimens. The two spare many patients in our institution the need for a major operative procedure to establish a secure tissue diagnosis. Immunohistochemistry does have a role to play but it is, at least in our opinion, clearly secondary to that of electron microscopy. The concept of replacing electron microscopy with a battery of immunostains has often been advocated as an economic measure, but this argument begins quickly to lose its weight as the number stains included in the battery is increased to cover the diagnostic possibilities. Giving consideration to the capriciousness of some of these stains, there exists with this also an increasing possibility of a spurious or misinterpreted result leading to an errant diagnosis.(ABSTRACT TRUNCATED AT 400 WORDS)

Carcinoma, Small Cell↗
PubMed1994

Variations in mitochondrial ultrastructure and dynamics observed by high resolution scanning electron microscopy (HRSEM).

Rat adrenal cortex was processed for high resolution scanning electron microscopy (HRSEM) to confirm tubular cristae, reported by transmission electron microscopy to be present in cortex mitochondria. Mitochondria in several other tissue and cell types were also observed and their ultrastructure confirmed by using three-dimensional, stereo, high resolution scanning electron microscopy. The mitochondria in rat and human hepatocytes as well as human skin fibroblasts grown in culture contained tubular cristae approximately 30 nanometers in diameter. The fibroblast mitochondria proved to be long, up to 46 micrometers and branching, as compared to those in liver which were spherical in shape. Cold adapted brown fat cells were packed with mitochondria, these containing plate or shelf-like cristae. Branched, rat striated muscle mitochondria were observed to curve around contractile protein filament bundles. The muscle mitochondrial cristae were found to be both tubular and plate-like, within the same mitochondrion. The ratio of tubular cristae to plate-like cristae varied considerably between muscle mitochondria. In order to use ultrastructural changes in mitochondria for differential diagnosis, and because 3D reconstruction of mitochondria based on transmission electron microscopy serial sections is severely limited in resolution, it is imperative to first develop a correct understanding of tissue specific, normal mitochondrial ultrastructure based on three-dimensional, HRSEM methods.

Adipose Tissue↗
PubMed1995

Quantitative analysis of chondroitin sulphate retention by tannic acid during preparation of specimens for electron microscopy.

The ability of tannic acid to enhance binding of glycosaminoglycans to purified collagen was analysed in an in vitro system using amino sugar analysis on an amino acid analyser, transmission electron microscopy, and scanning electron microscopy. Collagen was purified by digestion with trypsin, papain, and hyaluronidase. Purified collagen was incubated with hyaluronic acid or with chondroitin sulphate glycosaminoglycan and then treated with tannic acid. Tannic acid was found to enhance retention during preparation for electron microscopy of either of the glycosaminoglycans onto collagen fibres. The ability of tannic acid to enhance binding of collagen and glycosaminoglycans might explain, at least in part, its structural reinforcement effect on resected synovial joint-apposing surfaces during preparation for scanning electron microscopy.

Amino Sugars↗
PubMed1983

A simple technique for the visualization of whole mount cytoskeletons with transmission electron microscopy.

Examination of whole mount cells in the transmission electron microscope has been useful in studies of cellular architecture. The common technique is to grow cells directly on formvar-coated, gold grids for direct observation through a cell. We report a technique for obtaining whole mount preparations which requires neither fragile formvar films nor expensive, gold grids. Cells are grown on palladium-coated coverslips and processed for electron microscopy. The cells and the palladium substrate are separated from the coverslip. The cell-palladium complex is then picked up on copper grids as in thin section processing. We compare images of the cytoskeleton using our technique with images using previously described techniques and present preliminary observations of contracting cell models. Such contractions would tear formvar films if attempted on cells grown in the conventional manner for whole mount examination. Our technique allows cells to contract without tearing the underlying substrate.

Cytological Techniques↗
PubMed1976

Determination of the mass of viruses by quantitative electron microscopy.

The photometric method of quantitative determination of dry mass by electron microscopy has been applied to the study of various types of viruses: animal, plant, insect, and bacterial. The method is applicable to all viruses having a mass of 1 x 10-18g or greater. The molecular weight of viruses can be calculated from the mass value by multiplying it by Avogadro's number. In comparison to other methods of determining the molecular weight of viruses, sedimentation and diffusion, sedimentation equilibrium, light scattering, and electron microscopy counting, the method of quantitative electron microscopy is competitive. In some ways quantitative electron microscopy is superior to other methods for the determination of molecular weight: There is no limitation to the size of the virus, no experimental time involved and no concentration and purity of virus preparations required, and finally it is independent of the geometry of the virion. In one important aspect it is unique when compared to other methods; namely, it affords one the capacity to analyse individual virus particles.

Biophysical Phenomena↗
PubMed1983

Conductive staining of biological specimens for scanning electron microscopy with special reference to ligand-mediated osmium impregnation.

Biological specimens for scanning electron microscopy are made electron-conductive and non-charging by intense osmication with tannic acid, thiocarbohydrazide, hydrazine hydrate or other bifunctional ligands (or mordants). Such osmication also minimizes tissue shrinkage during dehydration and drying, reduces tissue damage during metal coating and scanning observations, and circumvents the use of a thick metal coating which hinders high resolution. Intense osmication when repeated eliminates the need for a metal coating and produces a well-contrasted scanning image with good resolution. Moreover, such osmication reduces tissue damage during ion implanting, enhances the depth of imaging, allows x-ray microanalysis without coated metals, and permits resin-embedded observations, as well as continuous or repeated microdissections in the scanning electron microscope without charging. Intense osmication forms a surface coat or thickens the cell membrane. This surface coat consists of electron dense deposits of 4-20 nm thickness. The hydrazine hydrate-osmium method can be used in the vapor phase. The vapor phase application has the advantage of staining even plastic samples for non-metal-coated scanning electron microscopy.

Animals↗
PubMed1981

Comparison of three electron microscopy techniques for the detection of human rotaviruses.

Rotavirus detection by direct electron microscopy was compared with direct and indirect immune electron microscopy techniques. The latter two approaches permitted the enumeration of 25 and 103 times more rotaviruses respectively, than direct electron microscopy. Also, 70% and 90% of the virus particles were aggregated by direct and indirect immune electron microscopy techniques respectively, thus facilitating their detection.

Antibodies, Viral↗
PubMedDate not supplied

Electron microscopy in the diagnosis of small round cell tumors of bone.

Small round cell tumors involving bone can present problems in differential diagnosis by light microscopy. In exploring the role of electron microscopy in this situation, seven small cell osteosarcomas and seven mesenchymal chondrosarcomas were examined by electron microscopy and compared with typical and atypical Ewing's sarcomas. There is much overlap in the ultrastructural features of these tumors, but electron microscopy is helpful to establish or confirm a diagnosis of typical Ewing's sarcoma and, if representative matrix is present, of small cell osteosarcoma.

Bone Neoplasms↗
PubMed1986

[Comparative scanning and transmission electron microscopy studies of the ependyma of the central canal in the spinal cord of primates. I. Electron optical image of the ependyma in the central canal of the spinal cord of the callithrix monkey (Callithrix jacchus, Linné 1758)].

The ependyma lining the central canal of the spinal cord of adult males and females monkey, Callithrix jacchus, was examined by scanning and transmission electron microscopy. The cross section of the lumen of the central canal are round, oval, or triangular. Light and dark ependymal cells, depending on the density of the cytoplasm, were found. The light ependymal cells are fewer than the dark cells. The ependyma cytoplasm contained numerous mitochondria, filamentous structures, one or more well-developed Golgi-complexes, vesicles of the smooth endoplasmic reticulum, ribosomes, lysosomes, multivesicular bodies, profiles of the rough endoplasmic reticulum, large osmophilic bodies, and microtubules. The nuclei of the ependyma cells usually have a simple, regular round or oval shape. They occupy a relatively large portion of the cell volume and lie in the central or mediobasal position. Some of the nuclei show deep invaginations into the karyoplasm. Most of the mitochondria occupy mainly the supranuclear portion of the apical cytoplasm. There are of the crista-typ. Ribosomes occur free in the cytoplasm, but some attached to the profiles of the rough endoplasmic reticulum or being arranged as polysomes. The filamentous structures are generally prominent cytoplasmic components and are distributed at the apical, lateral, or basal region of the ependymocytes. They are grouped into bundles and arranged in parallel arrays. Some of these bundles reach the plasmamembrane at the free lumina of the central canal, others take contact to the filamentous structures of the zonulae adherentes of the junctional complex below the free surface. The granular endoplasmic reticulum shows specializations. There profiles surrounding granular substances and widely distributed granulations in connection with the nuclear envelope. The functional significance of the deposition of these granulations is still unknown. The luminal surface of the ependymocytes bears many microvilli and cilia. The cilia are regularly arranged in cranio-caudal direction. Each cilium has the typical (9 + 2)-subfibres. The intercellular space at the surface of the ependymal layer shows a single zonula adherens or zonulae adherentes in the row. Tight junctions and gap junctions were not found in the material examined. Cell processes of liquor contacting neurons between adjacent ependyma cells, protruding into the lumen of the central canal, could be observed. The termination of these neurons contains accumulations of mitochondria in the central part, large amounts of vesicles, and small dense bodies. They have short microvilli and some stereocilia at the free surface.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1979

Immune labeling of certain strains of Actinomyces naeslundii and Actinomyces viscosus by fluorescence and electron microscopy.

A total of 12 well-characterized strains of Actinomyces viscosus and A. naeslundii grown on Trypticase soy agar plates supplemented with sheep erythrocytes were examined by light microscopy and transmission electron microscopy after treatment with appropriately labeled antisera to homologous and heterologous strains. Cells incubated with homologous rabbit antisera followed by fluorescein-isothiocyanate (FITC)-conjugated goat anti-rabbit immunoglobulin G (IgG) exhibited a completely smooth fluorescent outline in the case of A. naeslundii and and interrupted, irregular fluorescent outline in the case of human strains of A. viscosus. The different labeling patterns appeared to be related to the presence at the ultrastructural level of long, unevenly distributed strands of "fuzz" on the surface of human A. viscosus cells, whereas A. naeslundii cells had a narrower layer of fuzz, or more even thickness. The immunocoating reaction revealed homologous antibody binding to the irregular strands of fuzz on the surface of human A. viscosus cells, whereas homologous antisera to A. naeslundii coated A. naeslundii cells with a moderately electron-dense coating of antibody of even thickness. Human strains of A. viscosus incubated with heterologous antiserum to A. naeslundii followed by FITC-labeled goat anti-rabbit IgG exhibited a segmented fluorescent outline, which differed from that produced with homologous antisera. A. naeslundii incubated with heterologous rabbit antisera to human A. viscosus strains and FITC-labeled anti-rabbit IgG exhibited a completely smooth fluorescent outline similar to that produced with homologous antiserum. A. viscosus strains of hamster origin differed from A. viscosus strains of human origin by the absence of a surface fuzz and the comparatively smooth, even fluorescence produced by incubating these cells with homologous rabbit antiserum followed by FITC-labeled goat anti-rabbit IgG. Antiserum to a hamster strain did not cross-react with A. naeslundii or human strains of A. viscosus. Under the growth conditions of this experiment, ultrastructural features and labeling patterns with the indirect fluorescent technique may be useful in differentiating these serotypes from one another.

Actinomyces↗
PubMedDate not supplied

Fluorescence microscopy used in conjunction with horseradish peroxidase localization and electron microscopy for studying sympathetic nuclei of the rat spinal cord.

After application of horseradish peroxidase (HRP) to the transected cervical sympathetic trunk, labeled sympathetic neurons were seen in the intermediolateral nucleus (IML), central autonomic nucleus (CAN) and intercalated nucleus of rat spinal cords. As studied by glyoxylic acid-induced histofluorescence microscopy (FM), catecholaminergic (CA) terminals were most densely packed in the IML. There were 60 +/- 2 CA varicosities per 2,200 mu2 area in 20 mu thick sections through the IML. A combined FM/HRP method confirmed that preganglionic sympathetic neurons are heavily innervated by CA terminals. CA boutons were tagged with 5-hydroxydopamine for EM identification, and in the IML, 56% of CA boutons were seen to make synaptic contacts as compared to 60% in the CAN. It seems likely that virtually all CA boutons may form synapses but serial sections through boutons were not studied. It is inferred that preganglionic sympathetic neurons of both IML and CAN are served by CA inputs, and electron microscopy has revealed that most or all of these CA terminals transmit their signals via synapses; which may permit more precise structural sorting than non-synaptic transmission.

Adrenergic Fibers↗
PubMed1978

Low temperature scanning electron microscopy: a review.

Low temperature scanning electron microscopy is useful for morphological and analytical studies both in situations where low temperature techniques are used during specimen preparation and where low temperature stages are used for specimen examination and analysis. Examples are given of different low temperature specimen preparation techniques and how they may be applied to different types of specimen. There are still a number of problems associated with morphological identification in fully frozen-hydrated samples and it is important to carry out parallel studies using more conventional transmission electron microscopy and light microscopy preparation techniques. A number of criteria are presented, some or all of which may be used to establish the existence of the frozen-hydrated state.

Animals↗