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PubMed1983

Ultrastructural pathology of Crohn's disease: correlated transmission electron microscopy, scanning electron microscopy, and freeze fracture studies.

The mucosa of the ileum or colon from 16 patients with active Crohn's disease was studied using transmission electron microscopy of thin sections, scanning electron microscopy, and freeze fracture electron microscopy. Electron micrographs of diseased tissue, as well as of grossly normal resection margins, were studied. Correlations of mucosal disease, as seen with the various ultrastructural techniques, were made and compared to controls. Alterations of epithelial cell tight junctions in freeze fracture replicas correlated with a decrease in the number of pinpoint contacts between adjacent cells in transmission electron microscopy preparations. Gap junctions in Crohn's disease did not vary with respect to structure or number, when compared to controls. Mucosal microulcerations, affecting 1 to 6 cells, were observed in transmission and scanning electron microscopy preparations. Possible freeze fracture electron microscopy correlates of microulcers consisted of replicas of cells completely devoid of tight junctional structures. "Aphthoid" type ulcers were studied with the various ultrastructural tools and were remarkable for several abnormalities of villi bordering the aphthoid ulcers, as well as surrounding villi. Transmission, scanning, and freeze fracture electron microscopy each provide unique and confirming data that may be used to construct a model for the pathophysiology of Crohn's disease.

Colitis↗
PubMed1979

Morphological studies on the periostracum of the fresh-water mussel Amblema (Uniondae): light microscopy, transmission electron microscopy, and scanning electron microscopy.

The structure of the periostracum in the fresh-water mussel Amblema has been described using light microscopy, transmission elec;ron microscopy, and scanning electron microscopy. The structure and evolutive course of the periostracum was studied along its entire length, from the periostracal groove until it forms the tough outer covering of the shell. At least five structurally and functionally distinct regions were identified. In addition, the periostracum itself was seen to be a multilayered structure consisting of three major layers which are themselves subdivided into minor layers. From these morphological observations, a regulatory role for the various periostracal layers in mineral trapping, nucleation, and the subsequent formation of the prismatic and nacreous layers of the shell can be postulated.

Animals↗
PubMed1990

Correlative light microscopy, scanning electron microscopy, and transmission electron microscopy of osmium-macerated biological tissues.

A method facilitating correlation of light microscopic (LM), scanning electron microscopic (SEM) and transmission electron microscopic (TEM) images was developed. Rat kidney and heart were initially subjected to the osmium maceration procedure and then embedded in acrylic resin. Semithin sections of the tissue blocks were first provided for LM and then examined by SEM after resin removal. Furthermore, the ultrathin sections adjacent to the semithin sections were observed by TEM. The three-dimensional images of intracellular organelles provided an informative adjunct to LM and TEM.

Animals↗
PubMed1983

Scope of scanning electron microscopy, transmission electron microscopy and freeze fracture technique in diagnostic cytology of effusions.

A review of the literature on the application of scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to cytodiagnosis of neoplastic cells in effusion fluid reveals a lack of consensus as to which of the techniques is most beneficial to a cytologist for a satisfactory diagnosis. The purpose of this study is therefore to critically evaluate the relative merits of SEM, TEM and freeze-fracture (FF) techniques in enhancing the diagnostic "sensitivity" of ultrastructural cytology. Two cases of adenocarcinoma, one case of mesothelioma and one case of small cell undifferentiated carcinoma of the lung were studied in detail by SEM, TEM and FF techniques. A comparative analysis of the results demonstrates that SEM is a rapid and useful technique where the microvilli on the tumour cell surface is the most prominent feature especially in adenocarcinomas. The major handicap of SEM becomes evident in cases where the extreme variability of the surface morphology amongst tumour cells makes it difficult to establish a diagnosis with confidence. TEM along with light microscopy (LM) of toluidine blue sections gives us the most comprehensive method for cyto-diagnosis and is particularly useful in cases where LM cytology alone is inconclusive. It has the added advantage of revealing diagnostically significant nuclear and cytoplasmic features which are helpful in differentiating a benign from a malignant cell and in some instances the probable site of the primary neoplasm. FF technique, though not conducive to a routine diagnostic situation, is of benefit in demonstrating the organization of junctional complexes. From preliminary studies it appears that the tight junctions in adenocarcinoma are different from those seen in small cell carcinoma. In mesothelioma, gap junctions are invariably present, but they are not seen prominently in adenocarcinoma or small cell carcinoma. Thus in poorly differentiated tumours where LM and TEM are inconclusive, FF may prove useful. It is concluded that such coordinated studies in the future have the potential to better our understanding of the nature and behaviour of tumour cells in effusion fluid.

Adenocarcinoma↗
PubMed1985

Hürthle-cell lesions of the thyroid: a combined study using transmission electron microscopy, scanning electron microscopy, and immunocytochemistry.

Hürthle cell transformation found in 2 nodular goiters, 2 cases of Hashimoto's thyroiditis, 4 follicular adenomas, 3 follicular carcinomas, 2 papillary carcinomas and 1 anaplastic carcinoma were studied by transmission electron microscopy, scanning electron microscopy and immunocytochemistry. Ultrastructural features of Hürthle cells were identical in non-neoplastic and neoplastic lesions. Cells crammed with mitochondria, showing abnormalities in size, shape and content were prominent in most cases. The presence of distinct smooth-surfaced cells interspersed with cells with many microvilli is almost a pathognomonic scanning electron microscopic feature of benign and malignant Hürthle cell lesions. Most Hürthle cells stained positively for thyroglobulin in all cases, but no immunoreactivity for CEA and calcitonin was found.

Adenocarcinoma↗
PubMed1991

Historical perspective and current trends in emission microscopy, mirror electron microscopy and low-energy electron microscopy. An introduction to the proceedings of the Second International Symposium and Workshop on Emission microscopy and Related Techniques.

Emission microscopes and related instruments comprise a specialized class of electron microscopes that have in common an acceleration field in combination with the first stage of imaging (i.e., an immersion objective lens, also called a cathode lens or emission lens). These imaging techniques include photoelectron emission microscopy (PEEM or PEM), electron emission induced by heat, ions, or neutral particles, mirror electron microscopy (MEM), and low-energy electron microscopy (LEEM), among others. In these instruments the specimen is placed on a flat cathode or is the cathode itself. The low-energy electrons that are emitted, reflected, or backscattered from the specimen are first accelerated and then imaged by means of an electron lens system resembling that of a transmission electron microscope. The image is formed in a parallel mode in all of the above instruments, in contrast to the image in scanning electron microscopes, where the information is collected sequentially by scanning the specimen. A brief history and introduction to emission microscopy, MEM, and LEEM is presented as a background for the Proceedings of the Second International Symposium and Workshop on this subject, held in Seattle, Washington, August 16-17, 1990. Current trends in this field gleaned from the presentations at that meeting are discussed.

China↗
PubMed1982

Comparison of solid-phase immune electron microscopy, direct electron microscopy and enzyme-linked immunosorbent assay for detection of rotavirus in faecal samples.

One hundred and fifteen faecal samples from children with gastroenteritis have been examined by solid-phase immune electron microscopy (SPIEM), standard direct electron microscopy and enzyme-linked immunosorbent assay (ELISA). Diluted high-titered anti-(human) rotavirus serum, produced in rabbit by immunization with purified pooled rotaviruses from gastroenteritis patients, was used as antiserum in the immunological tests. The presence of rotavirus was demonstrated in 41 (36%) of the 115 samples by both SPIEM and ELISA. No false positive reactions were seen. By direct electron microscopy rotavirus was detected in 35 (30%) of the specimens. In addition, other gastroenteritis viruses like adeno-, astro- and calicivirus were visualized by this non-specific test in eight samples. The applicability of these three methods in routine diagnostic work is discussed.

Antigen-Antibody Reactions↗
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↗
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↗
PubMed1977

Colloidal gold, a useful marker for transmission and scanning electron microscopy.

Electron dense markers of a size suitable for transmission electron microscopy and scanning electron microscopy have been prepared with gold granules labeled with a monolayer of specific macromolecules. The optimum conditions for preparing the markers have been ascertained. The method is simple, rapid and seems to be general since gold granules have been labeled with polysaccharides and proteins. As homogeneous populations of gold granules having different sizes can be prepared, the method is also suitable for double marking experiments. The gold technique is illustrated by the localization of polysaccharides and glycoproteins on yeast cell walls and erythrocyte membranes by transmission electron microscopy and on yeast cells and intact erythrocytes by scanning electron microscopy. Good spatial resolution of the marker was achieved in all cases. The method is also suitable for marking thin sections. Spectrophotometric measurements were used to determine the number of gold granules adsorbed per cell.

Candida↗
PubMed1983

[Muscular changes caused by suction drainage. Scanning electron microscopy, transmission electron microscopy, light microscopy and morphometric studies of dorsal muscles of the rat (Rattus rattus) using drainage].

Changes in the dorsal muscles of 22 rats (Wistar), resulting from surgical drain, have been studied under different morphological aspects. The lumen of drains were obstructed after a short time. In the drains without using low pressure, erythrocytes and fibrin network were seen by scanning microscopical investigation. Patent muscle fibres could be analyzed in the lumen of drain using negative pressure. They prevent the functional drain system. By the effect of surgical drains the fibres of dorsal muscles had become swollen. This fact could be analyzed by morphometrical technique. Some changes in fibres have been demonstrated by histological methods.

Animals↗
PubMed1986

Pulmonary artery endothelial abnormalities in patients with congenital heart defects and pulmonary hypertension. A correlation of light with scanning electron microscopy and transmission electron microscopy.

Scanning electron microscopy and transmission electron microscopy were applied to lung biopsy specimens from patients with congenital heart defects, and pulmonary artery endothelium was analyzed for alterations in surface characteristics and intracytoplasmic composition which might reflect abnormal function. The patients were divided into four groups distinguished by increasing severity of pulmonary vascular changes on light microscopy graded both morphometrically and by the Heath-Edwards classification; group 1, normal vasculature or only abnormal extension of muscle into peripheral arteries; group 2, medial hypertrophy; group 3, medial hypertrophy +/- decreased artery number + intimal hyperplasia; group 4, decreased artery number + occlusive intimal hyperplasia. On scanning electron microscopy, the pulmonary artery endothelial surface in group 1 patients was "crinkled" or "corduroy-like", i.e., composed of narrow, even ridges; in groups 2 and 3, it was "cable-like", i.e., comprised of deep intertwined ridges; in group 4 it was "chenille" in texture, i.e., high ridges alternated with low, uneven, and twisted ones. There was significant increased density of surface microvilli in groups 2 and 3 patients when compared to groups 1 and 4 (p less than 0.05 for each comparison). On transmission electron microscopy pulmonary artery endothelial cells in groups 2 and 3 patients were also characterized by a significant increase in the volume density of rough endoplasmic reticulum (p less than 0.01) and microfilament bundles (p less than 0.05). The coarse endothelial surface characteristics associated with pulmonary vascular changes may result in abnormal interaction with blood elements and release of vasoactive substances. The increased microvilli, rough endoplasmic reticulum, and microfilament bundles in patients with moderate but not advanced arterial changes suggest a phase where increased endothelial metabolic function and alterations in the cytoskeleton may also contribute to heightened pulmonary vascular reactivity.

Actin Cytoskeleton↗