ISSN:1052-0295    

DOI:10.3109/10520299509108326

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

The present study shows that Langerhans cells can be differentiated from Interdigitating cells at the light microscopic level. Superficial lymph nodes and skin taken from necropsies and the lymph nodes of dermatopathic lymphadenopathy (DPL) were used for this experiment. Sections of lymph node and skin were embedded using the acetone, methyl benzoate and xylene (AMeX) method and dendritic cells were immunostained with anti S-100 protein antibody (S-100, and OKT-6 (CD1a) using the restaining method. Langerhans cells in the skin were positive for both CD1a and S-100. Dendritic cells positive for both CD1a and S-100, and dendritic cells positive for S-100, but not for CD1a were observed in superficial lymph nodes. In normal superficial lymph nodes, there were more interdigitating cells than Langerhans cells. The majority of the dendritic cells in the DPL were Langerhans cells. We conclude that the S-100 and CD1a positive cells are Langerhans cells, and the S-100 positive-CD1a negative cells are interdigitating cells.

ISSN:1052-0295    

DOI:10.3109/10520299509108327

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Fixation and staining of planaria can affect the interpretation of histopathological changes following their exposure to various agents. We assessed several fixation protocols with various stains in planaria to determine an optimal combination. Planaria were fixed in each of the following: 10% neutral buffered formalin, 2.5%, glutaraldehyde, Bouin's, Zenker's, 70% ethanol, and relaxant. In addition, planaria were fixed in relaxant and postfixed in each of the fixatives above. Paraffin embedded sections from each fixation protocol were stained with hematoxylin and eosin (H&E), toluidine blue, periodic acid-Schiff (PAS), or phosphotungstic acld-hematoxylin (PTAH). Relaxant fixed planaria were also stained with Steiner's, Holmes, trichrome, Giemsa, Grocott's methenamine silver (GMS) and antibodies for intermediate filaments (cytokeratin, vimentin and desmin). Relaxant and Zenker's gave the best fixation with minimal artifacts. Formalin, glutaraldehyde, and ethanol were unacceptable because they caused contortions of the body, crenation, and a darkly pigmented epidermis. Gastroderm could be differentiated from stroma best when stained with H&E, toluidine blue and PTAH. Other organ systems differentially stained included the epidermis, marginal adhesion gland, nervous tissue, and muscle. PAS, Steiner's, Holmes, trichrome and the intermediate filament stains were not useful for planaria staining. The most morphological information was obtained with relaxant fixative and a combination of sections stained with H&E and PTAH.

ISSN:1052-0295    

DOI:10.3109/10520299509108328

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Granule protein deficiencies in morphologically mature neutrophil cells of peripheral blood from human patients with acute myeloid leukemia was demonstrated using post-embedding immunocytochemistry. Abnormal immunoreactivity of granule proteins was detected in seven of nine patients. Decreased immunoreactivity patterns were found more for the primary granule markers elastase and myeloperozidase than for the secondary granule marker lactoferrin. Leukemias with a predominant myeloid component, in contrast to those with a predominant monocytoid component, had more neutrophil cells showing immunodeficiencies for one or more granule markers. The proportion of neutrophil cells showing immunodeficiencies varied greatly for each granule marker; more variation was obtained for elastase, lactoferrin and myeloperoxi-dase than for lysozyme, possibly because lysozyme is a marker for both granule types. In addition, no correlation could be found between any of the immunoreactivity deficiencies for the neutrophil granule glycoproteins elastase, lactoferrin, lysozyme and myeloperozidase and the abundance of a particular set of ultrastructural features in the circulating leukemic cells from any of the nine patients. Nonetheless, most of the immature myeloid cells from peripheral blood of leukemic patients showing neutrophil protein im-munoreactivity abnormalities in one or more granule markers often and randomly displayed one or more unusual ultrastructural features. The clinical and pathological significance of neutrophil granule protein deficiencies and the abundance of fibrillar structures in malignant myeloid cells presently is uncertain.

ISSN:1052-0295    

DOI:10.3109/10520299509108329

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Quantitative immunoelectron microscopy (QIEM) is dependent on the reliability of preparative techniques for both efficient immunolabeling and consistent quantitative results among series of immunostained sections. The present study compared Lowicryl K4M and Epon embedding after identical fixation and dehydration of rat somatotrophic secretory granules. Labeling intensity, diameter, roundness, uptake of uranyl acetate, and gray value were measured with computer assisted image analysis. Lowicryl-embedded granules showed the highest labeling densities after conventional fixation and Progressively Lowering Temperature (PLT) dehydration, but values were not consistent in a series of immunostained sections. A lower but more uniform level of immunostaining was seen in Eponembedded sections when tissue was cryofixed and physically dehydrated. Gray value measurements from micrographs from both embedding media confirmed the better contrast of Epon sections and the different reliefs of the granule surfaces. This study emphasizes the importance of complete comparisons of preparative techniques for QIEM for reliability and reproducibility.

ISSN:1052-0295    

DOI:10.3109/10520299509108330

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

A number of in situ hybridization protocols using digoxigenin or biotin labelled probes were assessed for viral nucleic acid detection in formalin fixed, paraffin embedded tissue. Single-step detection protocols for biotin labelled probes produced low sensitivity; however, enzyme based one-step detection protocols for digoxigenin probes produced high sensitivity for both RNA and DNA systems. For both probe types, multistep detection protocols produced equally high sensitivity. Use of an enhanced APAAP procedure for digoxigenin labelled probes acheived maximal sensitivity without use of biotin-streptavidin reactions. The sensitivity of nucleic acid detection obtained with a digoxigenin labelled probe is comparable to that obtained using biotin. Digoxigenin labelled probes for nucleic acid detection are recommended for tissues with endogenous biotin.

ISSN:1052-0295    

DOI:10.3109/10520299509108331

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

The effects of storing 2-hydroxy-ethyl methacrylate (HEMA) solutions for embedding tissues for light microscopy were studied using three commercially available HEMA embedding kits: Technovit 7100, Technovit 8100, and JB-4. These HEMA solutions were examined at various times of storage over a period of one year using a panel of physicochemical techniques including gas chromatography, tltration, viscosimetry, determination of the maximum polymerization temperature and the time required to reach the maximum temperature, and detection of degradation products of HEMA monomers by histochemical procedures. The quality of the resin blocks was examined by the observation of mini-folds in sections. Data obtained from these tests showed that the release of by-products as a result of the degradation of the HEMA monomer during storage of HEMA solutions does not occur. Development of cross-linking agents by transesterification of HEMA monomer was not detected either. Gradual decrease of the inhibitor concentration during storage proved to be the main cause of the reduction of shelf-life of HEMA solutions. Inconsistent tissue infiltration after storage may be due to decreased rates of tissue penetration as a result of HEMA chain lengthening. Guidelines for safe and economical handling of HEMA mixtures are given.

ISSN:1052-0295    

DOI:10.3109/10520299509108332

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor