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PubMed1995

Electron microscopy of the seminiferous epithelium in the triploid (ZZZ and ZZW) fowl, Gallus domesticus.

The ultrastructure of cells of the seminiferous epithelium is described in both triploid ZZZ and ZZW fowls, especially cytological aberrations influencing the development of morphologically abnormal spermatids and spermatozoa. Mitotic divisions in triploid fowls are similar to those in diploid fowls, but meiosis in both the ZZZ and ZZW triploid fowl is abnormal and many defective gametes are produced. The majority of spermatids have structural abnormalities and this is attributed to the aneuploidy expected from meiotic chromosome pairing in a triploid. Some spermatids were observed to possess an irregularly shaped head, 2 centriolar complexes, 2 acrosomes and 2 tails and this type differentiated into large-headed spermatoza. Such spermatids and spermatozoa are suggested to be near diploid due to their large nuclear size and 2 sets of organelles. Other spermatids, observed with only 1 centriolar complex, 1 acrosome and 1 tail, are suggested to be near haploid. Differentiation of these spermatids is similar to the process observed in diploid fowls except that there is a high percentage of abnormally shaped heads. The fact that morphologically normal spermatozoa were rare is consistent with the expectation that very few spermatozoa would be exactly haploid. In both ZZZ and ZZW triploid fowls, triple pairing of chromosomes is observed in synaptonemal complexes (SC) during meiosis and recombination nodules are distributed at random in the central regions of the triple pairing SC. In ZZW triploid fowls gamete formation is more severely affected than in ZZZ triploid fowls and it is suggested that many of the degenerative changes occurring in ZZW fowls could be attributed to those spermatids that possess a W sex chromosome, since normal male fowls are the homogametic sex (ZZ) and normal spermatids and spermatozoa contain only a Z sex chromosome.

Animals↗
PubMed1985

Use of immunosorbent electron microscopy for detection of rota- and hepatitis A virus in sucrose solutions.

Immunosorbent electron microscopy was used to demonstrate rotavirus in solutions of varying sucrose concentrations after 18, 42 and 66 h of incubation. About 50% of adsorption of virus particles to the grid was achieved after 18 h incubation and nearly 100% after 42 h when compared to trapping of virus from sucrose free solutions. Hepatitis A virus was purified in a 10-30% sucrose gradient and each fraction was examined by immunosorbent electron microscopy, direct electron microscopy, immune electron microscopy and radioimmunoassay. The sensitivities of immunosorbent electron microscopy and radioimmunoassay were essentially similar and considerably greater than direct electron microscopy and conventional immune electron microscopy.

Antibodies, Viral↗
PubMed1991

In defence of aldehyde-osmium fixation and critical-point drying for characterization of seed-storage fungi by scanning electron microscopy.

Low-temperature scanning electron microscopy cannot be used as a general approach when resolution of definitive surface features is required for fungal characterization, because of the production of a pervading exudate by some of the species, strains or varieties. As the resolution of the light microscope is obviously limiting, scanning electron microscopy remains the best microscopical mode. Results are presented for four species within the Aspergillus flavus group which indicate that minimal (if any) distortion or collapse can be achieved with concomitant preservation of surface morphology, when particular precautions are taken during fixation and critical-point drying. Details of the steps pertaining to preparation for scanning electron microscopy following critical-point drying that are considered collectively to contribute to the success of the approach are discussed.

Aldehydes↗
PubMed1984

A scanning and transmission electron microscopy study of jaagsiekte lesions.

A scanning electron microscopy (SEM) and transmission electron microscopy (TEM) study was made of lesions from acute, experimentally induced cases of jaagsiekte. In the SEM study tumour cells were easily identified by the abundant microvilli on their peripheral surface. The SEM study gave further insight into the development of lesions and the spatial relationship of cells involved in jaagsiekte. TEM revealed that the tumour cells were in a state of rapid protein synthesis and had many characteristics in common with other malignant cells.

Animals↗
PubMed1996

The osmium tetroxide-p-phenylenediamine procedure reveals the chromatid cores and kinetochores of meiotic chromosomes by light and electron microscopy.

We analyzed first-metaphase meiotic chromosomes of the grasshopper Chorthippus jucundus by two different methods, i.e., a silver impregnation technique and the osmium tetroxide-p-phenylenediamine (Os-PPD) procedure. The former was applied on squashed testes previously fixed in ethanol-acetic acid, whereas for Os-PPD the material was not subjected to any previous extraction treatment but was fixed in OsO4, treated with PPD, and embedded in Epon 812. Both techniques revealed chromatid cores and kinetochores regardless of the processing of the material (squashed or sectioned). Unstained Os-PPD sections were analyzed by light microscopy and transmission electron microscopy (TEM). The Os-PPD technique provided a high contrast of chromatid cores and kinetochores in relation to the chromatin, which revealed a low electron density. To determine the Os-PPD reaction mechanism, the PAS procedure, as well as scanning electron microscopy (SEM) backscattering and SEM X-ray microanalysis, was performed on sections. By use of the Os-PPD-PAS procedure, glycol groups formed by oxidation of osmium bound to aromatic substrates were detected in chromatid cores and kinetochores by brightfield and fluorescence microscopy. A high Z contrast was detected in these structures by backscattered electron imaging. SEM X-ray microanalysis showed osmium and phosphorus to be the main elements present on the chromatid cores. Taking into account the known reactivity of OsO4 and the present results, the possible participation of nucleic acids as well as proteins in the Os-PPD reaction mechanism and in the composition of chromatid cores and kinetochores is discussed.

Animals↗
PubMed2003

Object oriented database and electronic notebook for transmission electron microscopy.

As high-resolution biological transmission electron microscopy (TEM) has increased in popularity over recent years, the volume of data and number of projects underway has risen dramatically. A robust tool for effective data management is essential to efficiently process large data sets and extract maximum information from the available data. We present the Electron Microscopy Electronic Notebook (EMEN), a portable, object-oriented, web-based tool for TEM data archival and project management. EMEN has several unique features. First, the database is logically organized and annotated so multiple collaborators at different geographical locations can easily access and interpret the data without assistance. Second, the database was designed to provide flexibility to the user, so it can be used much as a lab notebook would be, while maintaining a structure suitable for data mining and direct interaction with data-processing software. Finally, as an object-oriented database, the database structure is dynamic and can be easily extended to incorporate information not defined in the original database specification.

Databases, Factual↗
PubMed1988

Cholangiovenous reflux pathways as defined by corrosion casting and scanning electron microscopy.

Using corrosion casting and scanning electron microscopy of the rat biliary tree, we investigated the site and size of the pathways that allow bacteria to reflux from bile to blood. Nonobstructed rat biliary trees were injected retrograde with methylmethacrylate resin at a constant rate of 0.04 ml/min to volumes of 40, 60, 80, 120, 160, and 200 microliters. The infusion pressure was monitored and a pressure-volume curve was constructed. After polymerization and corrosion in 30 percent potassium hydroxide, the casts were examined with scanning electron microscopy. In addition, to identify the size of the reflux pathways, ceramic particles of 150 A, 1.7 mu, or 10 mu were added to the resin, and the studies were repeated. Finally, intact livers with casted biliary trees were processed and studied by scanning electron microscopy without corrosion. Scanning electron microscopy demonstrated fine anatomic detail of the cholangiovenous reflux pathway. At 40 microliters (20 cm water pressure), normal biliary radicals were filled. Between 40 and 80 microliters (20 to 50 cm water pressure), the cast material refluxed from the bile ductules into the spaces of Mall and Disse and then into the hepatic sinusoids. Filling of sinusoids continued at volumes between 80 to 160 microliters, and filling of collecting veins was seen above 160 microliters. Particles of 1.7 mu and smaller readily refluxed, but there was no sinusoidal reflux of casting material that contained particles of 10 mu. Casting without corrosion showed that the liver parenchyma remained intact. There was no evidence of reflux across hepatocytes. This study shows that cholangiovenous reflux occurs directly from bile ductules through the spaces of Mall and Disse into the hepatic sinusoids. The pathways measure between 1.7 and 10 mu. Since this is the path of least resistance, it may be of greater importance in the reflux of bacteria and toxins than other high-resistance pathways, for example, biliary canaliculi, tight junctions, or hepatocytes.

Animals↗
PubMed1991

Detection of Norwalk-like virus and specific antibody by immune-electron microscopy with colloidal gold immune complexes.

Direct electron-microscopy (DEM), immune electron microscopy (IEM) and four different procedures of immune electron microscopy with colloidal gold immune complexes were evaluated for the detection of Norwalk-like virus and specific antibody. A solid-phase immune electron microscopy with colloidal gold immune complexes-triple layer method (SPIEMGIC-TLM) is developed for screening patients' specimens for the detection of Norwalk-like virus and its specific antibody. The method demonstrates low non-specific background labelling and is simple, sensitive and easy to perform. A quadruple layer method (SPIEMGIC-QLM), which is a modification of the triple layer method, has been established by adding a cross-linking anti-IgG layer to amplify the reaction and to provide a more sensitive test which is suitable for screening monoclonal antibodies prepared against 32-34-nm Norwalk-like virus isolated in our laboratory.

Antibodies, Monoclonal↗
PubMed1980

Scanning electron microscopy - some methodological considerations.

Although scanning electron microscopy has been used in arterial research for more than a decade, methodological standards are still subject to dispute. In this presentation, our experience with different methods to prepare arterial tissue is described. Our efforts have been aiming at optimizing the reproducibility of our preparations. The recommended procedure involves fixation at "physiological" arterial pressure in Karnovsky's fixative for a short period, post-fixation by immersion, drying of the tissue with the critical point method and covering it with gold in a sputtering device. The effects of various deviations from this procedure are described. After preparation with the recommended procedure a resolution of 200-400 A has been obtained. General characteristics of the arterial endothelium are concurrent with those described in studies using light microscopy or transmission electron microscopy.

Animals↗
PubMed1983

Scanning electron microscopy of intestinal metaplasia of the human stomach.

Biopsy samples of gastric mucosa were obtained at endoscopy from patients suspected of having intestinal metaplasia. These samples were prepared for light microscopy and scanning electron microscopy. Observations of the luminal surface of the gastric mucosa in the scanning electron microscope revealed distinct topographic differences in the intestinalized and nonintestinalized regions. The latter area showed features consistent with those described in chronic gastritis, i.e., the exaggeration of the "cobblestone relief" appearance of the luminal surface of the stomach and the presence of numerous short, stubby, bulbous microvilli which project from the free border of the surface mucous cells toward the lumen of the stomach. The intestinalized region revealed a plush microvillous (striated) border periodically interrupted by apical openings of goblet cells. The scanning electron microscope is a useful research aid for examination of cell membrane expressions of the surface cells of the gastric mucosa not totally appreciated by light microscopy and transmission electron microscopy.

Endoscopy↗