ISSN:1052-0295    

DOI:10.3109/10520299309105568

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

A technique was developed for sectioning fresh red spruce foliage (Picea rubensSarg.) for use in fluorescence microscopy. This allowed rapid examination of mesophyll in 3–5 mm needle sections. Healthy, ozone treated and cold stressed needles were examined to assess the utility of this technique for early detection of damage. Healthy mesophyll cells fluoresced bright red, while injured cells fluoresced yellow-green in ozone treated needles, and yellow-orange in frozen needles. Shifts in fluorescence wavelengths may be useful for early detection of injury to mesophyll before it is evident by standard light or electron microscopy.

ISSN:1052-0295    

DOI:10.3109/10520299309105569

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Numerous studies have described the F-actin cytoskeleton; however, little information relevant to C-actin is available. The actin pools of bovine aortic endothelial cells were examined using in situ and in vitro conditions and fluorescent probes for G-(deoxyribonuclease I.0.3μM) or F-actin (phalloidin, 0.2μM). Cells in situ displayed a diffuse G-actin distribution, while F-actin was concentrated in the cell periphery and in fine stress fibers that traversed some cells. Cells of subconfluent or just confluent cultures demonstrated intense fluorescence, with many F-actin stress fibers. Postconfluent cultures resembled the condition in situ; peripheral F-actin was prominent, traversing actin stress fibers were greatly reduced and fluorescent intensity was diminished. Postconfluency had little influence on G-actin. with only an enhancement in the intensity of G-actin punctate fluorescence. When post-confluent cultures were incubated with cytochalasin D (15 min; 10--4M), F-actin networks were disrupted and actin punctate and diffuse fluorescence increased. G-actin fluorescence was not altered by the incubation. Although its unstructured nature may account for the minor changes observed, the stability of the G-actin pool in the presence of notable F-actin modulations suggested that filamentous actin was the key constituent involved in these actin cytoskeletal alterations. A separate finding illustrated that the concomitant use of actin probes with image enhancement and fluorescent microscopy could reveal simultaneously the G- and F-actin pools within the same cell.

ISSN:1052-0295    

DOI:10.3109/10520299309105570

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

To facilitate cell kinetics studies of brain tumors labeled with thymidine analogs, we developed a new method to identify nuclei labeled sequentially with bromodeoxyuridine (BUdR) and iododeoxyuridine (IUdR) by double staining with immunogold-silver and alkaline phosphatase. Patients received an intraoperative infusion of BUdR: excised tumor specimens were immediately labeled with IUdR in vitro. fixed with 70% alcohol, embedded in paraffin, and cut into 6pmsections. The sections were incubated first with BR-3. a monoclonal antibody that recognizes only BUdR, and then with IU-4. a monoclonal antibody that recognizes both BUdR and IUdR: sections were counterstained with hematoxylin to identify unlabeled nuclei. Nuclei labeled only with IUdR stained red, whereas those labeled with BUdR or with both BUdR and IUdR stained black against a red background: unlabeled nuclei stained blue. This method was the most efficient differential staining technique to identify nuclei labeled only with IUdR and those labeled with BUdR among unlabeled nuclei.

ISSN:1052-0295    

DOI:10.3109/10520299309105571

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

The usefulness of thionin for staining cartilage sections embedded in glycol meth-acrylate (GMA) and the effect of decalcification on cartilage sections embedded in paraffin and GMA were assessed. Short decalcification periods using 5% formic acid or 10% EDTA did not influence the staining properties or the morphology of cartilage matrix and chondrocytes. The standard stain safranin O-fast green for differential staining of cartilage was used as control in these experiments. Prolonged exposure of safranin P stained sections to fast green resulted in disappearance of the safranin O stained matrix, thereby hampering the quantitative measurement of negatively charged glycosaminoglycans (GAG). Thionin stained evenly throughout all cartilage layers, independent of the staining times. In contrast to safranin 0, thionin did not show meta-chromasia in nondehydrated cartilage sections, which made it more suitable for assessing cartilage quality in GMA embedded cartilage. To evaluate the selectivity of thionin staining in cartilage, chondroitinase ABC and trypsin digestions were carried out. Thionin staining was prevented by these enzymes in the territorial matrix, representing the interlacunar network and the chondrocyte capsule. Staining with thionin of the interterritorial matrix was only slightly reduced, possibly representing keratan sulfate and hyaluronic acid in cartilage of elderly patients. Comparison of thionin stained GMA embedded cartilage with safranin O stained paraffin embedded sections showed significant similarity in optical densitometry, indicative of the specificity of thionin bound to negatively charged GAG in cartilage. In GMA embedded cartilage morphology was relatively intact compared to paraffin embedded sections due to less shrinkage of chondrocytes and the interlacunar network.

ISSN:1052-0295    

DOI:10.3109/10520299309105572

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

The neutral red and methylene blue in vitro cytotoxicity assays were compared under a variety of conditions using normal human ovarian epithelial cells to determine whether either assay is superior for studying cell growth. The results were standardized against a DNA spectrofluorometric assay. Although the assays were equivalent in reflecting cell number, each has specific advantages: while neutral red discriminates between viable and dead cells, the methylene blue assay is more sensitive and easier to perform.

ISSN:1052-0295    

DOI:10.3109/10520299309105573

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

During extraction of hemicellulose and noncellulose polysaccharides from plant histological slices of plants the morphological integrity of the slices may be disturbed if the reagents extracting hemicellulose and noncellulose polysaccharides are rinsed away by water. Removing the reagents by delicate suction rather than removing the glass tray from the staining dish, followed by desiccation and a rinse in 95% ethanol terminates the treatment without altering the morphology of the tissues.

ISSN:1052-0295    

DOI:10.3109/10520299309105574

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

A new antiroll device has been developed to replace the antiroll guide plate for cryostat wax sectioning. With this device, a continuous ribbon of 3–4μm sections can be obtained. The sections are flat, uncreased. and compression is reduced to a minimum.

ISSN:1052-0295    

DOI:10.3109/10520299309105575

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Psammoma bodies and small calcifications are frequently seen in a wide variety of tissues. These deposits of calcium salts render tissues difficult to section. Conventional decalcification alters the tissues; consequently it is not advisable to decalcify tissues containing only small calcium deposits. A simple and rapid method to remove small amounts of calcium salts using citric acid alone is described. This method does not alter the antigenic properties of the substances studied.

ISSN:1052-0295    

DOI:10.3109/10520299309105576

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

A simple and reliable technique is described for in situ excision of distended Samples of greater omentum from small laboratory animals. The omental bursa is distended by injecting whipped hen egg white. Filter paper frames then are applied to the selected areas of distended omentum and samples of omental membrane are excised together with the filter paper frames. This sampling technique yields undamaged materials suitable for various research purposes.

ISSN:1052-0295    

DOI:10.3109/10520299309105577

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor