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At least 253 records · Page 14Linked to original sources
PubMed1975

Wide-field electron microscopy. A rapid method for the study of histologic material that provides a bridge between light and electron microscopy.

Wide fields of tissue can be rapidly examined by electron microscopy by use of Formvar films for the support of ultrathin sections on slot grids. The intervention of the grid bars of conventional mesh grids is avoided, and a continuous micrograph of the specimen at scanning magnifications can be obtained. Enough material is sublimed from the section and the supporting film by deliberate exposure to the electron beam to permit one to obtain an image with good contrast. This method of examination, which takes in all about two hours, permits examination of an extensive area of tissue in relation both to its topography at low magnifications and to its ultrastructural detail, and accordingly adds to electron microscopy a dimension characteristic of the lower powers of the light microscope. It offers the histopathologist the option of using micrographs taken during the scanning survey of a tissue to detect regions that can be readily re-examined at high magnification in the same ultrathin section.

Animals↗
PubMed2002

Computer-assisted simulation of high-voltage electron microscopy using serial images recorded by conventional transmission electron microscopy.

Here we describe a computer-assisted method which, based on conventional transmission electron microscopy, renders simulated high-voltage electron micrographs. We perform arithmetic minimum filtering on stacks of aligned serial transmission electron microscopic images. In this way, the structural information of the separate images is fused into one compound image that highlights organization patterns otherwise easily overlooked or impossible to comprehend. Our method, like high-voltage electron microscopy, offers the possibility to build stereo-pairs for high-resolution three-dimensional analysis of tissue layers 1-2 microm thick. The use of background elimination and the development of a depth enhancement routine improved the three-dimensional effect and facilitated the analysis of the interior of objects. As an example, we use the method to display the distribution of axonal organelles at nodes of Ranvier and the shape and contents of a highly branched hippocampal dendritic spine.

Animals↗
PubMed1992

Low-energy electron microscopy (LEEM) and mirror electron microscopy (MEM) of biological specimens: preliminary results with a novel beam separating system.

Low-energy electron microscopy (LEEM) and mirror electron microscopy (MEM) utilize a parallel beam of slow-moving electrons backscattered from the specimen surface to form an image. If the electrons strike the surface an LEEM image is produced and if they are turned back just before reaching the surface an MEM image results. The applications thus far have been in surface physics. In the present study, applications of LEEM and MEM in the biological sciences are discussed. The preliminary results demonstrate the feasibility of forming images of uncoated cultured cells and cellular components using electrons in the threshold region (i.e. 0-10 V). The results also constitute a successful test of a novel beam-separating system for LEEM and MEM.

Animals↗
PubMed1994

A new method to correlate acoustic spectroscopic microscopy (30 MHz) and light microscopy.

A powerful new method is used to investigate the correlation between light microscopic and acoustic properties of biological tissues. Specimens of liver were sectioned into successive slices, 250 micrometers and 10 micrometers thick. The thick sections were investigated acoustically, the thin sections by means of light microscopy. Markers that could be detected and located, both optically and acoustically, were used to find and reconstruct corresponding regions in the acoustic and optical sections (2.5 x 2.5 mm). Parameter images were reconstructed from the sections investigated acoustically. The acoustic parameters were attenuation at 30 MHz, the slope of the attenuation spectrum (between 10 and 50 MHz), backscattering at 30 MHz, the slope of the backscattering spectrum (between 10 and 50 MHz) and the local ultrasound velocity. Acoustic images were obtained in the frequency range from 10 to 50 MHz, yielding a lateral resolution of about 50 micrometers. The sections for light microscopy were stained according to the Goldner trichrome staining technique. The histological composition was determined quantitatively, using digital image segmentation techniques. The percentage of collagen-rich fibrous tissue, luminal structure and interstitial spaces, and the number of nuclei were calculated for regions of 250 x 250 micrometers. These histological features were correlated with the acoustic parameters obtained from the corresponding regions in adjacent sections. It was thus possible to find the histological components responsible for acoustic parameters.

Animals↗
PubMed1990

A new light microscopy classification of spermatogenesis in the bull and the boar applicable to tissues processed for electron microscopy.

A new light microscopy classification of spermatogenesis in the bull and the boar was formulated to be used in conjunction with electron microscopy. It uses information available from all cell types and allows for very small portions of testicular epithelium at any orientation to be evaluated. The classification devised, being simple in outline, allows for the natural overlap of cell types. Furthermore, it emphasises the direct comparability of the morphological events occurring during spermatogenesis in the bull and the boar.

Animals↗
PubMed1986

[Sites of aluminum accumulation in bone marrow: study using electron microscopy, ionic microscopy and X-ray microanalysis].

Two methods of analytical microscopy have been used to study the distribution of aluminum in bone marrow of rats intoxicated by aluminum gluconate. Images of the distribution of aluminum in a field of 250 microns in diameter were obtained by analytical ion microscopy. They show that this element was concentrated in spots, associated with iron or alone, in the cytoplasm of some cells. Electron Probe Microanalysis (EPMA) has shown that aluminum concentration occurred in cells of the reticulo-endothelial system, principally in the reticular cells of erythroblastic islets. In cells of the reticuloendothelial system, aluminum was observed in intracytoplasmic organelles having ultrastructural characteristics of lysosomes or phagolysosomes. In these organelles, aluminum is always associated with phosphorus and sometimes with iron. No cytoplasmic or nuclear aluminum accumulation was detected in any other variety of bone marrow cells. The consequences of the selective accumulation of aluminum in the cytoplasm of reticular cells of erythroblastic islets for the maturation of erythrocytes are discussed.

Aluminum↗
PubMedDate not supplied

Unwarping confocal microscopy images of bee brains by nonrigid registration to a magnetic resonance microscopy image.

Confocal microscopy (CM) is a powerful image acquisition technique that is well established in many biological applications. It provides 3-D acquisition with high spatial resolution and can acquire several different channels of complementary image information. Due to the specimen extraction and preparation process, however, the shapes of imaged objects may differ considerably from their in vivo appearance. Magnetic resonance microscopy (MRM) is an evolving variant of magnetic resonance imaging, which achieves microscopic resolutions using a high magnetic field and strong magnetic gradients. Compared to CM imaging, MRM allows for in situ imaging and is virtually free of geometrical distortions. We propose to combine the advantages of both methods by unwarping CM images using a MRM reference image. Our method incorporates a sequence of image processing operators applied to the MRM image, followed by a two-stage intensity-based registration to compute a nonrigid coordinate transformation between the CM images and the MRM image. We present results obtained using CM images from the brains of 20 honey bees and a MRM image of an in situ bee brain.

Animals↗
PubMed1989

Response of microbial adhesives and biofilm matrix polymers to chemical treatments as determined by interference reflection microscopy and light section microscopy.

The polymers involved in the adhesion of Pseudomonas fluorescens H2S to solid surfaces were investigated to determine whether differences between cell surface adhesives and biofilm matrix polymers could be detected. Two optical techniques, i.e., interference reflection microscopy (IRM) and light section microscopy (LSM), were used to compare the responses of the two types of polymer to treatment with electrolytes, dimethyl sulfoxide (DMSO), and Tween 20. To evaluate initial adhesive polymers, P. fluorescens H2S cells were allowed to attach to glass cover slip surfaces and were immediately examined with IRM, and their response to chemical solutions was tested. With IRM, changes in cell-substratum separation distance between 0 and ca. 100 nm are detectable as changes in relative light intensity of the image; a contraction of the polymer would be detected as a darkening of the image, whereas expansion would appear as image brightening. To evaluate the intercellular polymer matrix in biofilms, 3-day-old biofilms were exposed to similar solutions, and the resultant change in biofilm thickness was measured with LSM, which measures film thicknesses between 10 and 1,000 microns. The initial adhesive and biofilm polymers were similar in that both appeared to contract when treated with electrolytes and to expand when treated with Tween 20. However, with DMSO treatment, the initial adhesive polymer appeared to contract, whereas there was no change in thickness of the biofilm polymer. These results indicate that both polymers bear acidic groups and thus act electrostatically with cations and are able to enter into hydrophobic interactions.(ABSTRACT TRUNCATED AT 250 WORDS)

Adhesins, Bacterial↗
PubMed1981

Comparison of direct electron microscopy, immune electron microscopy, and rotavirus enzyme-linked immunosorbent assay for detection of gastroenteritis viruses in children.

An approximate 10% suspension in water of the first available stool sample from 411 infants and young children with acute gastroenteritis was examined by electron microscopy (EM) after 2 min of negative staining. This procedure enabled the detection of 88% of the 199 rotavirus infections, all of the 22 adenovirus infections, and 47% of the 15 approximately 27-nm virus infections ultimately detected by a combination of techniques, including immune electron microscopy (IEM) and rotavirus enzyme-linked immunosorbent assay (ELISA). Of the 204 infections detected by direct EM of stools, 76% were detected within 2 min of viewing, and 94% were detected within 6 min of viewing. Type 1 and type 2 rotavirus particles were visualized with approximately equal efficiency, although type 2 rotavirus infections were more common. Rectal swab preparations were clearly inferior to stool preparations for the detection of virus infection by direct EM. IEM examination was required for efficient visualization of viruses in rectal swab specimens. ELISA was the most sensitive method for the detection of rotaviruses; with this method, all infections in which rotavirus particles were visualized by EM or IEM were detected. However, 73% of the 1,834 specimens which were presumptively positive for rotavirus by conventional indirect ELISA proved to be falsely positive on the basis of EM, IEM, blocking ELISA, confirmatory ELISA, or a combination of these methods. False-positive rotavirus ELISA reactions apparently were eliminated when fecal specimens were tested in a modified confirmatory ELISA with a lower dilution of rotavirus-negative (pre-immunization) than rotavirus-positive (post-immunization) capture antibody from the same animal.

Acute Disease↗
PubMed1983

Comparison of solid-phase immune electron microscopy by use of protein A with direct electron microscopy and enzyme-linked immunosorbent assay for detection of rotavirus in stool.

A total of 525 stool specimens collected during 1 year were examined for the presence of rotavirus by direct electron microscopy (EM), enzyme-linked immunosorbent assay (ELISA), and a solid-phase immune electron microscope method (SPIEM) utilizing protein A-coated grids for anchoring of specific viral antisera. Rotavirus was seen in 187 specimens; SPIEM detected 183 (97.8%), whereas direct EM and ELISA detected 161 (86%) and 166 (88.7%), respectively. No false-positive reactions were seen by ELISA. The sensitivity of the methods was evaluated by coded investigation of a dilution series of a positive sample, with a negative fecal specimen as diluent. SPIEM was approximately 30 times more sensitive than direct EM and 10 times more sensitive than ELISA. A study was done to compare the elapsed time for recognition of rotavirus by SPIEM and EM in 25 randomly selected positive specimens. All virus-positive specimens were detected within 2 min by SPIEM, whereas up to 9 min was required for direct EM. SPIEM with protein A is a highly sensitive method, useful for rapid detection of viruses in clinical specimens. Due to the direct visualization of virus particles by electron microscopy, there is no requirement for monospecific antisera for the method.

Enzyme-Linked Immunosorbent Assay↗
PubMed1996

Modified technique to recover microsporidian spores in sodium acetate-acetic acid-formalin-fixed fecal samples by light microscopy and correlation with transmission electron microscopy.

Microsporidia are an emerging cause of significant disease, particularly in the immunocompromised host. Until recently, the diagnosis of enteric infections has required invasive sampling, the use of expensive technology, and considerable technological expertise. The purpose of the present study was to examine three modifications to the processing of fecal specimens for light microscopy (LM) examination for microsporidian spores: the use of pretreatment with potassium hydroxide, modified centrifugation conditions, and a modified staining technique. A sodium acetate-acetic acid-formalin-fixed fecal sample containing numerous microsporidian spores confirmed to be positive by transmission electron microscopy (TEM) was used in all studies performed. A simulation of a heavy to lightly infected individual was used. The results of LM were correlated with those of TEM. Duplicate smears were stained with Weber's modified trichrome and Giemsa (GS) stains. The stained slides were randomized and examined blindly by LM at x 625 and x 1,250 magnifications. A portion of the dilutions after centrifugation were fixed for TEM. The Weber modified trichrome stain performance rating was higher than the Giemsa stain rating because of ease of interpretation, and material stained with Weber modified trichrome stain required less examination time at a lower magnification. The number of positive smears and the quantity of spores detected were significantly higher following pretreatment of the sample with KOH. TEM was positive only when numerous spores were present, but the quality of the photomicrographs was superior after pretreatment with KOH. Pretreatment of sodium acetate-acetic acid-formalin-fixed fecal samples with 10% KOH and then a 5-min centrifugation time and staining with Weber modified trichrome stain provide for the excellent recovery of microsporidia in the routine diagnostic parasitology laboratory.

AIDS-Related Opportunistic Infections↗
PubMed2004

Fibered confocal fluorescence microscopy (Cell-viZio) facilitates extended imaging in the field of microcirculation. A comparison with intravital microscopy.

This study investigated the capability of fibered confocal fluorescence microscopy (FCFM) to provide in vivo microvascular observations. FCFM is specifically designed for in vivo in situ observation thanks to a probe composed of a fiber bundle and micro-optics having a diameter as small as 650 microm. In the first part of the study, we compared the main characteristics of FCFM with those of intravital fluorescence microscopy (IFM). A mouse cremaster preparation was used as a common basis to allow for imaging with both modalities. We discussed the feasibility of obtaining quantitative measurements usually provided by IFM in the context of FCFM: morphometry, capillary permeability, functional capillary density, vasoconstriction and dilation effects. In addition, the possibility to visualize fluorescent red blood cells or leukocytes was also evaluated. Phototoxicity issues and limitations of FCFM were also discussed. We showed that FCFM allows observations and measurements usually provided by IFM and that the real-time capability of the system, as well as the flexibility and small diameter of the optical probe enable micro-invasiveness and can extend imaging capabilities for in vivo in situ observations when compared to IFM.

Animals↗
PubMed1990

Electron microscopy, immune electron microscopy, enzyme immunoassay and immunofluorescent evaluation of rotaviruses isolated from individual calves and piglets.

Rotaviruses were isolated in primary bovine foetal kidney cells from 3 of 13 (23.1%) tested suckling calves and from 3 of 14 (21.4%) suckling piglets both suffering of acute diarrhoea, as confirmed by serological tests. The presence of rotaviral antigen in culture supernatant was revealed by double sandwich enzyme immunoassay (EIA) and direct immunofluorescence (IF). Transmission electron microscopy (TEM) and immune electron microscopy (IEM) showed particles with an average diameter of 70 nm. The strains adapted to continuous pig kidney cells PK-15 have lost their reactivity in EIA, although TEM revealed the presence of rotavirions.

Animals↗
PubMed1988

[Distribution of lymph vessels in the plica vocalis of the human. A light microscopy, enzyme histochemistry and electron microscopy study].

Results of studies on the presence of lymphatic capillaries in the plica vocalis are shown, using improved methods of preparation. Immediately after laryngectomy, followed by immediate fixation of the non-pathological plicae vocalis, large lymphatic capillaries can be easily identified in the specimen. Lymphatic capillaries, which cannot be seen in conventional light microscopy, are now histochemically demonstrated with a marker of 5' nucleotidase. These histochemical results are confirmed by electron microscopy. Using a combination of both methods, a different quantity of subepithelial lymphatic capillaries can be visualised over the whole length and width of the plica vocalis. Lymphatic capillaries below the stratified squamous epithelium in the region of the anterior commissure grow gradually more numerous up to the folds of the arytenoid cartilage. Hence, in contrast to previous studies, the superficial lymphatic capillary system of the laryngeal mucosae is not separated into a supraglottic and subglottic lymphatic net of the vocal cord.

5'-Nucleotidase↗
PubMed1985

Identification of a subpopulation of primary granules in human neutrophils based upon maturation and distribution. Study by transmission electron microscopy cytochemistry and high voltage electron microscopy of whole cell preparations.

Two main classes of primary granules have been identified in human neutrophils by peroxidase cytochemistry and electron microscopy, spherical granules and elongated granules, containing crystals with well-defined periodicity. In the present study, we report another subpopulation of primary granules, distinguishable from the other primary granules by their small size, strong peroxidase reactivity under selective incubation conditions with diaminobenzidine, stage of neutrophil maturation at which they appear, and distinctive morphological characteristics. Early promyelocytes, reacted with diaminobenzidine before fixation, demonstrate no reactivity in the large primary granules, but strong peroxidase activity in the endoplasmic reticulum and Golgi apparatus. At the end of the promyelocyte stage of maturation, when the endoplasmic reticulum decreases in peroxidase reactivity, small, round or elongated granules (100 to 200 nm) are observed, reacting strongly for peroxidase. These small granules persist during maturation. The data suggest that these small granules are packaged after the large primary granules and before the secondary granules. All three classes of primary granules exhibit peroxidase activity when fixed prior to incubation in diaminobenzidine at neutral or alkaline pH. Distinctive morphological characteristics of the small primary granules are observed in circulating neutrophils. The small granules are arranged in chains or clusters primarily at the cell margin or uropod in moving cells and are aligned along the axis of cell polarity. This granule association is more evident in adherent neutrophils, particularly after short incubation with phorbol myristate acetate. Chains consisting of as many as 17 small granules are observed. Stereo high voltage electron microscopy of whole-mount preparations of adherent neutrophils reveals chains of small granules apparently interconnected by microtrabeculae. Thus, the small granules observed in thin sections do not represent a transverse section of elongated granules, and the interconnection of granules by microtrabeculae may determine the arrangement of granules. These data support the existence of subpopulations of primary granules which contain distinct forms of myeloperoxidase.

Bone Marrow↗
PubMed1988

The dento-gingival junction as seen with light microscopy and scanning electron microscopy.

The purpose of this paper is to review the anatomical relationship of the Dento-Gingival Junction as seen in the human dentition. The junction is described under light microscopy and then reviewed as seen in the SEM with the author's unpublished findings. The authors' material was derived from extracted human teeth with remaining marginal gingival tissue. The specimens were fixed with 2% glutaraldehyde in 0.15M sodium cacodylate buffer (pH 7.2) for 24 h. The specimens were then washed and freeze-fractured in Freon 113 using liquid nitrogen. Afterwards they were processed by freeze-drying or CPD methods, coated with gold, and placed in the scanning electron microscope (SEM) for viewing. These specimens demonstrated the presence of numerous Sharpey's fibers at the cemental surface. A large number of fibrils intermingled with the fibers to produce a dense mass of tissue. Junctional epithelium, with the adjacent homogeneous dental cuticle was demonstrated. Plaque deposits on the tooth surface extended to a cell-free zone. Morphological detail viewed with SEM and light microscopy are compared.

Freeze Fracturing↗
PubMed1983

[Bone structure in aseptic necrosis of the femoral head by light microscopy, contact microradiography, and fluorescent microscopy --comparing with osteoarthritis and rapidly destructive coxarthrosis].

In order to obtain further information of the pathological changes and bone structure of aseptic necrosis (AN), the present author carried out histological investigation on 20 femoral heads removed by prosthesis or total hip replacement from 18 patients. Both decalcified and undecalcified sections were investigated by microscopy, contact microradiography (CMR) and fluorescent microscopy. Schemata of individual cases were drawn to know the relationship between femoral head deformity and bone reactivity. Moreover, 19 femoral heads with secondary osteoarthritis (OA) removed from 18 patients and 7 with rapidly destructive coxarthrosis (RDC) from 6 patients were similarly investigated to compare with AN. The following results were obtained; Histologically, there are four different areas in AN. From the top of the femoral head, the lesion consisted of (1) subchondral necrosis, (2) granulation tissue, (3) an area of variable bone formation, and (4) normal bone tissue. As the advancement of the head deformity, fibrous component in the granulation increases and seems to divide the femoral head into necrotic and living areas, which by fibrous as well as enchondral ossifications become more prominent in the edge of the demarcation. As for the bone formation, four different types of bone tissues are discernible by CMR; (1) low calcified appositional bone and its absorption, (2) low calcified, appositional bone on wide trabeculae, (3) low calcified appositional bone on narrow trabeculae, and (4) irregularly calcified woven bone. Although during the comparatively early stage, the bone formation is seen in almost all of the living area, as the advancement of the head deformity, its activity decreases and results in overt head deformity to resemble that of OA which would be caused by mechanical factor. Increase of fibrous component in the granulation along with decrease of the bone formation would reduce the repairing activity of the femoral head. In the femoral heads with OA, a wedge-shaped area of bone formation is formed of which peak is located on the medial thickened cortex. In the femoral heads with RDC, a large amount of woven bone with irregular calcification is found in the bone formation area, in the upper part of a femoral head.

Adult↗