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PubMed1994

Changes in the distribution of F-actin in the fission yeast Schizosaccharomyces pombe by arresting growth in distilled water: correlative studies with fluorescence and electron microscopy.

Freeze-substitution electron microscopy of Schizosaccharomyces pombe cells starved in distilled water was conducted to define ultrastructural counterparts of actin visualized by fluorescence microscopy using rhodamine-conjugated phalloidin (Rh-ph). Starvation in distilled water caused remarkable changes in actin distribution and ultrastructural changes in S. pombe. Fluorescence microscopy of the starved cells showed that the dots of actin at the growing ends became thick actin cables via an enlarged patched form of actin. These changes were reversible, and growth-arrested cells resumed their original pattern of actin distribution upon return to growth medium. Electron microscopy of starved cells showed bundles of thin filaments and clusters of filamentous balls in the cytoplasm, which corresponded to the actin cables and enlarged actin dots, respectively, as seen by fluorescence microscopy. Vesicles polarized at the growing cell ends were dispersed in the cytoplasm by distilled water treatment, indicating that actin organization plays a role in directing vesicle location.

Actin Cytoskeleton↗

PubMed1979

Visualization of intrarenal catecholamine-containing elements: fluorescence histochemistry and electron microscopy.

Fluorescence histochemistry and electron microscopy were used to study catecholamine-containing elements in rat and cat kidneys. There were fluorescent nerve plexuses associated with the arterial system of the renal cortex from which nerves extended into the medulla forming dense networks in the vascular bundles of the outer part of the medulla. Also, there were small cells found singly and in clusters which emitted the blue fluorescence characteristic of catecholamine-containing cells. These cells were associated with the periarterial nerve plexuses and were frequently encountered in the cat medullary vascular bundles. Electron microscopy of the cat renal medulla revealed processes with the structure of typical adrenergic terminals and other processes that resemble small intensely fluorescent (SIF) cell processes.

Adrenergic Fibers↗

PubMed1985

Electron microscopy of body fluids.

Transmission electron microscopy and scanning electron microscopy are complementary techniques; the former is useful for examining the fine internal structural detail of tissue and cell surfaces and the latter is useful for studying three-dimensional configurations. The methods and findings of transmission and scanning electron microscopic study of effusions are summarized, and their practical application in routine clinical work is assessed.

Adenocarcinoma↗

PubMed2003

Extending energy-filtered transmission electron microscopy (EFTEM) into three dimensions using electron tomography.

The length scales on which materials microstructures are being formed, grown, and even designed are becoming increasingly small and increasingly three-dimensional. For such complex structures two-dimensional transmission electron microscopy (TEM) analysis is often inadequate and occasionally misleading. One approach to this problem is the modification of electron tomography techniques, developed for structural biology, for use in materials science. Energy-Filtered (EF) TEM elemental distribution images approximate to true projections of structure, and, as such, can be used to reconstruct the three-dimensional distribution of chemical species. A sample holder has been modified to allow the high tilt (+/-60 degrees ) required for tomography and a semiautomatic acquisition script designed to manage energy-loss acquisition. Tilt series data sets have been acquired from two widely different experimental systems, Cr carbides in 316 stainless steel and magnetite nanocrystals in magnetotactic bacteria, demonstrating single- and multiple-element tomography. It is shown that both elemental maps and jump-ratio images are suitable for reconstruction, despite the effects of diffraction contrast in the former and thickness changes in the latter. It is concluded that the image contrast, signal, and signal-to-noise ratio (SNR) are key to the achievable reconstruction quality and, as such, the technique may be of limited value for high energy loss/small inelastic cross section edges.

Electrons↗

PubMed2003

Detection of characteristic signals from as-doped (<1 at.%) regions of silicon by transmission electron microscopy and convergent-beam electron diffraction.

Characteristic signals were detected from As-doped (< 1 at.%) regions of silicon by dark-field transmission electron microscopy and convergent-beam electron diffraction. A slight intensity increase was observed in 220 dark-field images, which may be explained by an increase of scattering amplitude due to the As doping. The doped region showed a much higher intensity in 004 dark-field images. The characteristic high intensity was observed for specimens with As concentrations of about 0.09-0.8 at.%. Convergent-beam electron diffraction patterns obtained from the As-doped region showed a characteristic rocking curve for 004 reflection. These characteristics should originate from incoherent elastically scattered electrons due to a static lattice distortion around the doped As atoms. The observed characteristics in dark-field images and rocking curves of the 004 reflection should be a good probe not only for investigating the concentration of doped atoms in Si lattice, but also for the amount of impurity and/or point defects in other crystalline materials.

Journal Article↗

PubMed1997

Spatial distribution of dihydropyridine receptors in the plasma membrane of guinea pig cardiac myocytes investigated by correlative confocal microscopy and label-fracture electron microscopy.

Excitation-contraction coupling in cardiac muscle is thought to depend fundamentally on the spatial organization of sarcolemmal dihydropyridine receptors (L-type calcium channels) in relation to ryanodine receptors (calcium-release channels of the sarcoplasmic reticulum). In the present study, we have investigated the distribution of dihydropyridine receptors in the guinea pig myocyte plasma membrane by correlative immunoconfocal microscopy and label-fracture electron microscopy. Label-fracture, a method in freeze-fracture cytochemistry, permits immunogold localization of cell surface proteins in en face membrane views. Taken together, results from confocal microscopy and label-fracture replicas suggest that, in the peripheral plasma membrane, calcium channels are organized predominantly in the form of clusters. Confocal microscopy also suggests a similar organization in the transverse tubules. It is hypothesized that these clusters may lie adjacent to junctional sarcoplasmic reticulum, permitting the close coupling of influx of calcium through plasma membrane calcium channels to trigger release of calcium from the intracellular stores, as part of the mechanism of calcium-induced calcium release.

Animals↗

PubMed2005

Maxillary sinus augmentation using a synthetic cell-binding peptide: a histologic and transmission electron microscopy case study in man.

PepGen P-15 is a combination natural anorganic bovine-derived hydroxyapatite matrix coupled with a synthetic cell-binding peptide (P-15). This material has improved bone formation in periodontal osseous defects and bone regenerative procedures. There were 3 specimens retrieved 18 months after a sinus lifting procedure using PepGen P-15. These specimens were treated to be observed under light microscopy and transmission electron microscopy. Light microscopy showed that most of the particles were surrounded by newly formed bone. In some areas, osteoid matrix was present. No acute inflammatory infiltrate was present. In transmission electron microscopy, all phases of bone formation (i.e., osteoid matrix, woven bone, and lamellar bone) were observed in the newly formed bone around the biomaterial particles. In some regions, this newly formed bone seemed to present interdigitations connecting to or entering into the particle surface. To our knowledge, this is the first report presenting data on transmission electron microscopy of PepGen P-15 used in a sinus augmentation procedure in man. Our results confirm previous reports on the clinical effectiveness of this material.

Alveolar Ridge Augmentation↗

PubMed1993

Macrocyclization of polysaccharides visualized by electron microscopy.

Topological features of the polysaccharides schizophyllan, l-carrageenan and gellan gum were studied using electron microscopy. Electron micrographs of schizophyllan not subjected to any thermal or solvent composition history destabilizing the triple helix, show stiff, linear chains consistent with the structure being triple helical and with contour length proportional to the molecular weight in solution. A blend of linear, cyclic and hairpin topologies and higher molecular weight clusters were observed after renaturation, i.e. return to conditions favouring the triple helical structure, from solvent conditions dissociating the triple helix. Electron micrographs of l-carrageenan in salt-free solution reveal linear extended structures. Addition of 0.15 M LiI to the solution before preparation for electron microscopy, i.e. salt conditions that favour ordering but not gelation, yields a large fraction of cyclic structures with circumference of different lengths. Likewise, adding KCl to aqueous gellan gum changes their appearance from dispersed polymers to suprastrands with several associated chains. Macrocyclic species can also be observed in gellan gum after the addition of a gel-promoting salt. The tendency to form macrocyclic structures in competition with intermolecular aggregates is determined by the three factors: (1) chain stiffness relative to overall length; (2) parallel or antiparallel alignment of interacting chain segments; and (3) polymer concentration. The present study indicates that electron microscopy provides information about the topology adopted by polysaccharides.

Carbohydrate Conformation↗

PubMed1992

[Cell fine structures observed by scanning electron microscopy].

The scanning electron microscope (SEM) provides vivid seemingly three dimensional images which are easier to understand for us than transmission electron microscopic images. For this point of view scanning electron microscopy is advantageous in morphological researches of cell fine structures. Nevertheless, there were few studies in this field, because SEM had much lower resolution than transmission electron microscope (TEM) and because there was no adequate method to reveal intracellular structures. In recent years, however, the resolution of SEM has been markedly improved and the specimen preparation techniques have also advanced. In this paper, some of our preparation technique for revealing cell surface structures or intracellular structures, in particular, osmium-DMSO-osmium method, and the results observed by these methods were described. 1) Nucleus. The nucleus was wrapped with a nuclear envelope that consisted of two membranes enclosing a narrow space. On the surface of the envelope many nuclear pores were observed. 2) Endoplasmic reticulum (ER). Rough ER consisted of flattened cisternae, arranged in parallel. The surface were studded with many ribosomes which were often arranged spirally to form polysomes. Smooth ER consisted of tubules. 3) Golgi complex. a) The Golgi stacks were all linked by anastomosing. b) Connection between Golgi stacks and rough ER was often observed. c) Cisternae in a Golgi stack were connected each other. 4) Mitochondria. The mitochondrion was bounded by 2 sheets of unit membrane and the inner membrane projected into the interior of the organelles to make mitochondrial cristae.

Animals↗

PubMed2005

Use of light, scanning electron microscopy and bioassays to evaluate parasitism by entomopathogenic fungi of the red scale insect of palms (Phoenicococcus marlatti Ckll., 1899).

We have evaluated the parasitism of the red scale insect of the date palm (Phoenicococcus marlatti) by entomopathogenic fungi, using light microscopy (LM), scanning electron microscopy (SEM) and low temperature scanning electron microscopy (LTSEM). Beauveria bassiana, Lecanicillium dimorphum and Lecanicillium cf. psalliotae, were inoculated directly on the scale insects or on insect infested plant material. We found that L. dimorphum and L. cf. psalliotae developed on plant material and on scale insects, making infection structures. B. bassiana was a bad colonizer of date palm leaves (Phoenix dactylifera L.) and did not parasite the scale insects.

Cocos↗

PubMed1980

The demonstration of bacteria on and within the stratum corneum using scanning electron microscopy.

Using scanning electron microscopy it has been possible to demonstrate the location of bacteria on and within the stratum corneum of the human foot. Biopsies taken either by sectioning or by removing stratum corneum with cyanoacrylate ester adhesive were examined using a Jeol JSM-T20 scanning electron microscope. Bacteria could be seen easily on specimens from skin which had been occluded to increase the number of bacteria present. On the surface, bacteria were scattered widely in small colonies (usually containing less than ten bacteria). Although bacteria could be seen around the orifice of sweat ducts they did not preferentially favour these sites. Within the stratum corneum, bacteria could be found as relatively large colonies but these were usually associated with sweat ducts or the underside of the furrows in the skin surface. This study suggests that, in normal skin, bacteria are able to colonize both the surface and the depths of the stratum corneum.

Bacteria↗

PubMed2005

Histogenesis of human renal cell carcinoma by using electron microscopy and immunohistochemical techniques.

Electron microscopy and immunohistochemical techniques are powerful tools for the determination of tissue origin. Both techniques have been used in the current experiment for histogenesis of renal cell carcinoma. Fifty kidney tumors were subjected to immunohistochemical detection for intermediate filaments cytokeratin and vimentin, which are normally expressed in epithelial tissue and mesenchymal tissues, respectively. Twenty cases of the above were examined by electron microscopy for detection of ultrastructure features. From each kidney, two specimens were taken, one from the diseased area and another far from it to serve as a control. Immunohistochemical study revealed in cases of renal cell carcinoma, cytokeratin and vimentin were expressed alone in 44% of cases, and 40% of cases, respectively. Twelve percent of cases were coexpressed with both cytokeratin and vimentin. Electron microscopic study of diseased specimens revealed the expression of desmosomes which was observed in almost all tumor specimens. The expression of the vimentin in some cases either alone or with cytokeratin was interpreted as a change in the characters of some tumor cells which indicates the need for additional techniques in such cases to get the proper interpretation. The prevalence of the expression of cytokeratin and the persistence existence of desmosomes indicate the epithelial origin of the tumor. This data is very beneficial for determination of line of therapy and follow up of the patients. The results confirm the power of combined use of both immunohistochemistry and electron microscopy in the field of histogenesis.

Carcinoma, Renal Cell↗

PubMed1994

The loci of mineral in turkey leg tendon as seen by atomic force microscope and electron microscopy.

Transmission electron micrographs of fully mineralized turkey leg tendon in cross-section show the ultrastructure to be more complex than has been previously described. The mineral is divided into two regions. Needlelike-appearing crystallites fill the extrafibrillar volume whereas only platelike crystallites are found within the fibrils. When the specimen is tilted through a large angle, some of the needlelike-appearing crystallites are replaced by platelets, suggesting that the needlelike crystallites are platelets viewed on edge. If so, these platelets have their broad face roughly parallel to the fibril surface and thereby the fibril axis, where the intrafibrillar platelets are steeply inclined to the fibril axis. The projection of the intrafibrillar platelets is perpendicular to the fibril axis. The extrafibrillar volume is at least 60% of the total, the fibrils occupying 40%. More of the mineral appears to be extrafibrillar than within the fibrils. Micrographs of the mineralized tendon in thickness show both needlelike-appearing and platelet crystallites. Stereoscopic views show that the needlelike-appearing crystallites do not have a preferred orientation. From the two-dimensional Fourier transform of a selected area of the cross-sectional image, the platelike crystallites have an average dimension of 58 nm. The needlelike-appearing crystallites have an average thickness of 7 nm. The maximum length is at least 90 nm. Atomic force microscopy (AFM) of unstained, unmineralized turkey leg tendon shows collagen fibrils very much like shadow replicas of collagen in electron micrographs. AFM images of the mineralized tendon show only an occasional fibril. Mineral crystallites are not visible. Because the collagen is within the fibrils, the extrafibrillar mineral must be embedded in noncollagenous organic matter. When the tissue is demineralized, the collagen fibrils are exposed. The structure as revealed by the two modalities is a composite material in which each component is itself a composite. Determination of the properties of the mineralized tendon from the properties of its elements is more difficult than considering the tendon to be just mineral-filled collagen.

Animals↗

PubMed2005

Characterization of a peg-like terminal NOR structure with light microscopy and high-resolution scanning electron microscopy.

An atypical peg-like terminal constriction ("peg") on metaphase chromosomes of the plant genus Oziroë could be identified as a nucleolus organizing region (NOR) by detecting 45S rDNA with correlative light microscopy (LM) and scanning electron microscopy (SEM) in situ hybridization (ISH). Using high-resolution 3D analytical SEM, the architecture and DNA distribution of the peg-like NOR were characterized as typical for chromosomes, albeit with significantly smaller chromomeres. ISH procedure was improved for SEM concerning signal localization, labeling efficiency, and structural preservation, allowing 3D SEM analysis of the peg-like NOR structure and rDNA distribution for the first time. It could be shown that implementation of FluoroNanogold markers is an attractive tool that allows efficient immunodection in both LM and SEM. A model is proposed for the peg structure and its mode of condensation.

Cell Cycle↗

PubMed1985

The location of surface antigens of Bordetella pertussis by immuno-electron microscopy.

Immuno-electron microscopy using colloidal gold-tagged monoclonal antibodies (McAbs) has been used to detect antigens on the surface of Bordetella pertussis cells. McAbs to serotype-specific agglutinogens 2 and 3 labelled fimbriae in a serotype-specific manner. An attempt was made to determine whether individual fimbriae of serotype 1.2.3 cells bear both antigens 2 and 3. In double labelling experiments, individual cells were found to label with McAbs to antigen 2 (3 nm gold) and to antigen 3 (15 nm gold). Many fimbriae labelled with only one of these reagents, but there were instances where both labels could have been attached to the same fimbria. No fimbrial labelling was obtained with McAb to agglutinogen 1. Gold-tagged McAbs to filamentous haemagglutinin (FHA) labelled neither the fimbriae nor the surface of B. pertussis cells. Unfixed cells on electron microscope grids appeared to shed FHA readily. After fixation, a small proportion of cells bore aggregates of FHA which labelled specifically with anti-FHA McAb-gold. Results with one McAb to lymphocytosis promoting factor indicated that this antigen may be more intimately associated with the surface of B. pertussis than is FHA.

Antibodies, Monoclonal↗

PubMedDate not supplied

The continuing value of electron microscopy in surgical pathology.

For decades, transmission electron microscopy has played a valuable diagnostic role in surgical pathology. The continuing importance of electron microscopy, however, can be debated, given the major advances that have occurred in immunohistochemistry and other techniques. Electron microscopy retains excellent educational potential and broad research applicability, and it continues to be a necessity for the evaluation of a small subset of surgical pathology cases, such as renal biopsies and cilia specimens. The real controversy, then, centers on the contribution of electron microscopy in the evaluation of neoplasms. The opinion of many experts indicates that electron microscopy is still vital in the diagnostic assessment of some neoplasms, and that both electron microscopy and immunohistochemistry are more powerful when viewed as complementary rather than competitive techniques. For electron microscopy to be used to its potential, however, electron microscopists must function effectively as consultants. When optimally applied, electron microscopy remains an essential diagnostic tool.

Costs and Cost Analysis↗

PubMed2001

Free-floating cryostat sections for immunoelectron microscopy: Bridging the gap from light to electron microscopy.

Frozen skin sections are routinely used for light microscopic immunohistochemical study of the skin basement membrane zone for two reasons: some skin basement membrane zone proteins are labile to routine chemical fixation, and skin is not amenable to vibratome sectioning. However, inherent limitations of conventional frozen sections, including compromised morphology and a requirement for glass slide-mounting, usually limit immunohistochemical study to the light microscopy level. In the present study, we introduce use of unfixed, free-floating cryostat sections for characterization of immunolocalizations of selected skin basement membrane proteins at both the light and electron microscopy level. The new procedure employs free-floating cryostat sections that can be processed as routine tissue specimens and can be subjected to a variety of special staining procedures including immunohistochemistry. Especially useful is the ease of progressive processing of the same tissue specimen from light microscopy to electron microscopy. In this regard, the method renders itself useful when results of immunolabeling experiments need to be elucidated quickly at histological and ultrastructural levels as required for diagnostic and accelerated investigative strategies.

Antigens, CD↗

PubMed1984

Detection of monocytes and evaluation of their role in human glomerular pathology: the contribution of electron microscopy.

The potentialities of electron microscopy in detecting monocytes in human glomeruli are discussed and its reliability is compared with that of other techniques (such as enzyme histochemistry and immunohistochemistry) having the same purposes. The major advantages of electron microscopy are: the same material can be used for both monocyte detection and conventional diagnostic purposes; its high resolution power allows for information on the site of monocyte infiltration and clarification of some functions of the latter (such as the phagocytic activity on immune complexes or their participation to the basement membrane damage). Nevertheless, the suggested technique also has some disadvantages, in particular it permits to investigate only a small number of glomeruli and it creates difficulties in the identification of nonstimulated monocytes.

Glomerulonephritis↗