摘要:

Cells in the S-phase of the cell cycle can be identified in tissue sections by im-munohistochemical localization of the thymidine analogue bromodeoxyuridine (BrdU). Generally, a single counterstain is used to visualize the underlying tissue; however, interpretation of morphologic detail is often difficult. We have utilized BrdU to localize proliferating cells in myocardium exposed to angiogenic mitogens. To facilitate identification of labelled nuclei in the context of infarcted vs. viable myocardium, BrdU immunohistochemistry was followed by a modified Mas-son trichrome stain. The time of exposure to the counterstains and the wash protocol were re-revised, permitting clear identification of the labelled brown nuclei against a background of red viable myocardium vs. blue infarct. The combined technique also provides color contrast suitable for computer-based image analysis.

ISSN:1052-0295    

DOI:10.3109/10520299209110032

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

Gallyas technique was modified by a direct application of Schiff's reagent after physical development, resulting in distinctive staining of amyloid deposits in argyrophilic structures. With this modified method, senile plaques in Alzheimer's disease are clearer. This method is easy to perform and suitable for routine neuropathol-ogical examination.

ISSN:1052-0295    

DOI:10.3109/10520299209110033

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

The utility of fluorescence microscopy for studying development of fern spores is investigated. Changes in the fluorescence characteristics during the developmental stages of fern sporangia can be attributed to the changes in the chemical composition of the cell wall. Bright blue autofluorescence of the spores indicated the presence of sporopollenin. The sporan-gial walls and the spores autofluoresced yellow under blue light excitation. Fluorescence microscopy is a useful addition to light, scanning, and transmission electron microscopy because living specimens can be studied owing to their fluorescence properties.

ISSN:1052-0295    

DOI:10.3109/10520299209110034

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

The technique of in situ hybridization to both meiotic and mitotic chromosomes ofRumex acetosais described. Differences in the efficiency of signal detection were observed between the two types of material. The implications of these results for in situ hybridization to other plant species are explored.

ISSN:1052-0295    

DOI:10.3109/10520299209110035

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

We have performed a computer assisted image analysis evaluation of the effects of two preparation protocols on morphological variables and immunolabeling density of secretory granules using rat adenohypophysis as a model. Glutaraldehyde (GA) fixation for 2 hr and chemical dehydration was compared with short GA fixation (15 min) followed by cryofixation and freeze-drying (CF-FD). The 2 hr GA fixed specimens showed spherical nuclei and secretory granules regardless of their size, contrasting with the more irregularly shaped nuclei and secretory granules after short GA-CF-FD. In the latter group of specimens a correlation could be found between smaller nuclear areas and more irregular shapes. The differences in morphological variables between the two preparation protocols might be due to a better protein stabilization and a reduced collapse of macromolecules after the 2 hr GA fixation, strengthened by the chemical dehydration. The specific immunolabeling with antiserum to growth hormone was much greater but more varied after short time GA-CF-FD than after long GA fixation. Epon surface topography over the granule area also differed: smooth after short time GA-CF-FD and furrowed after long GA fixation. Our results, demonstrating important differences in morphological parameters and immunolabeling density between two common preparation protocols, seem critical for more reliable interpretation in quantitative immunoelectron microscopy. We also emphasize the need for computer assisted image analysis and measurements in immunoelectron microscopy to ensure objective evaluations.

ISSN:1052-0295    

DOI:10.3109/10520299209110036

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

A variant Golgi technique was developed that consisted of substituting osmium tetroxide with formaldehyde as the initial fixative in intracardiac perfusion, along with the addition of glacial acetic acid to the chromating fluid. This procedure avoids disposal of dangerous waste substances into the environment. Other advantages include 1) reduction of cost, danger to lab workers, and risk of disruption of the tissue slices during their handling by eliminating the osmium tetroxide, 2) clear tissue background, 3) greater quantity of impregnated neurons than in the classical procedure, with distinct morphological details easily identified even in gross sections and 4) reduction in processing time.

ISSN:1052-0295    

DOI:10.3109/10520299209110037

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

Age-related changes in cell proliferation kinetics of the mandibular condyle were studied in Sprague-Dawley rats using bromodeoxyuridine (BrdUrd) immunohistochemistry in decalcified, paraffin-embedded tissues. Intraperitoneal injection of 40 mg/kg BrdUrd was given 1 hr before animal sacrifice. Continuous perfusion of EDTA solution via the left ventricle shortened the decalcification time. Immunohistochemical staining was performed with monoclonal anti-BrdUrd antibody. BrdUrd labeled cells, i.e., the S-phase cells, were clearly visible with well-preserved cytological detail. Their nuclei exhibited homogeneously stained granules. The labeling index in the intermediate zone of the condyle decreased with increasing age of the animals. This method is useful for evaluating physiological and pathological changes of the rat mandibular condyle as well as other bones and joints.

ISSN:1052-0295    

DOI:10.3109/10520299209110038

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

A fairly simple method for calibrating microdensitometric histochemical assays is described. The method is based on paired biochemical and histochemical assays on single freeze-dried skeletal muscle fibers which differ widely in their properties. As an example, the method is applied to investigate the validity of the periodic acid-Schiff (PAS) reaction for the microdensitometric estimation of glycogen content. Some problems that may interfere with the calibration are discussed.

ISSN:1052-0295    

DOI:10.3109/10520299209110039

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

Elytra of the bessbeetle,Odontotaenius disjunctuswere thin sectioned after embedding in epoxy resin. Sections were cut with a diamond saw, ground to the desired thickness on a rotary grinder and polished. Tearing and distortion were reduced when compared to knife-cut sections of heavily sclerotized cuticle.

ISSN:1052-0295    

DOI:10.3109/10520299209110040

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

A simple method for handling electron microscope grids is described here. While assuring their identification and safety, this method provides improved handling, temporary storage, and identification of grids bearing ultra-thin sections, and in addition, provides a novel method for preparing bulk samples. Grids are attached at their edges to the weakly adhesive surface of a“Post-it”note pad which sits in a petri dish. The grids are safely immobilized on the pad, classified based on their location, and identified by convenient pad notation. Grid manipulation and identification are simplified using this device, which is easily assembled from readily available and inexpensive materials.

ISSN:1052-0295    

DOI:10.3109/10520299209110041

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor