ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.167

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractWe present a detailed protocol forIn Situhybridization of messenger RNA molecules in brain and other tissues, using35S-labeled single-stranded RNA transcribed from commercially available plasmid vectors. These vectors contain promoter sequences for specific synthesis of RNA, and polylinker regions that will accept cDNA inserts for virtually any target molecule of interest. The protocol is optimized for studies that include concomitant irnmunohistochemical and cytoarchitectonic evaluations, and it is sufficiently detailed for the molecular biologist unfamiliar with histologic technique or the histologist inexperienced in molecular biological methods. We have used it with consistant results forIn Situhybridization studies using probes to more than twenty different mRNAs; and for several of those we have obtained results showing quantitative differences in experimentally altered mRNA levels. Included are details of perfusion fixation, tissue sectioning and preparation, probe synthesis, hybridization reaction, and signal detection. The introduction and annotated comments include rationale for basic procedures and the purpose of various specific steps in the hybridization histochemistry protocol. (The J HistotechnoL12:169, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.169

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractPinocytotic vesicles are common organelles in cells of many different tissues. In the electron microscope, these vesicles appear as two types, coated and smooth. Coated vesicles have attracted recent attention with regard to the molecules involved in their formation and biochemical properties. Much less is known about smooth endocytic vesicles. In this study, we examined some of the properties of morphologically smooth vesicles in type I pneumocytes with immunological probes to coated vesicle protein components in a system where the smooth vesicle number is specifically increased by nitrogen dioxide (NO2). Using an indirect immunoperoxidase labeling technique with affinity purified anticalmodulin, anti-clathrinlight chains, and anti-clathrin-heavy chains, we were able to localize antigens on smooth vesicles that are commonly seen associated with coated vesicles by transmission electron microscopy of ultrathin sections. The labeled antigens were found on or around the vesicle membrane. In addition, results obtained with the listed primary antibodies in conjunction with an avidin-biotin-ferritin technique on thin sections embedded in Lowicryl confirmed the immunoperoxidase data. The three antigens were associated with smooth or uncoated pinocytotic vesicles. The data suggest that molecules reportedly associated with coated vesicles may also exist on smooth vesicles. It is hypothesized that some-of the molecular components that participate in the formation of coated vesicles may be involved with the formation of other vesicle types. (The J Histotechnol12:185, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.185

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractThe ultrastructure of the mouse adrenal cortex was compared after fixation either by immersion in or by perfusion with buffered glutaraldehyde. The most striking feature of the perfusion fixed cortex was the patency of the capillaries that, by comparison, were collapsed in immersion fixed cortex. In addition, the osmotically sensitive mitochondria and endoplasmic reticulum were swollen and damaged only in immersion fixed tissue. In perfusion fixed adrenal cortices, the profiles of the mitrochondria in all three major cortical zones ranged from ovoid to elongated; their cristae were tubular or tubulosaccular (not lamelliform) and arranged in a parallel fashion. Zonal variations in mitochondria1 ultrastructure in immersion fixed cortex may reflect fixation artifacts and are not suitable criteria for distinguishing the cortical zones. In perfusion fixed adrenal cortex, the intensity of the osmium staining of lipid-filled cytoplasmic vesicles was markedly reduced, particularlyin the iona fasciculata, which by comparison was heavily stained by osmium in the immersion fixed cortex, indicating retention of osmiophilic lipids. This perfusion protocol was developed for use in conjunction with electron microscopic autoradiography for mapping radiolabeled lipoprotein uptake in mouse adrenal glands. (J Histotechnol12:195, 1989.)

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.195

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractIncreased interest in quantitation of the histopathological changes in a variety of neurological disorders (including neurodegenerative disease such as Alzheimer's disease) continues in an attempt to develop specific clinical-histopathological correlations. Most previous efforts at quantitation have used paraffin embedded sections of brain tissue, although plastic embedded sections have recently become a preferable alternative because they provide greatly reduced tissue shrinkage and distortion during processing, and greater clarity and improved resolution to the tissue sections. We have developed techniques for glycol methacrylate embedding and sectioning of brain tissue blocks on a standard histology laboratory microtome. In addition, we have modified routine diagnostic and investigational neurohistological stains for use in glycol methacrylate embedded brain sections, including hematoxylin and eosin, modified Bielschowsky stain, Jamarri silver technique, Einarson's Nissl stain, gallocyanin-Darrow red myelin stain, and the thioflavine-S-hematoxylin stain. The use of plastic embedded sections with appropriate stains will permit critical histopathological evaluation of nervous system tissue from patients with a variety of neurological disorders. (J Histotechnol12:201, 1989)

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.201

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractParaffin embedded sections fixed in 10% formalin were stained in Shandon Instant Hematoxylin, differentiated in acid alcohol, blued in dilute sodium acetate, and counterstained with a concentrated eosin-multichrome stain. This simple procedure produced consistent results with deep blue nuclei and bright counterstaining. (J Histotechnol12:210, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.210

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractCreutzfeldt-Jakob disease (CJD) resembles two other diseases, kuru (found in New Guinea tribesmen) and scrapie (found in sheep). CJD is a slow progressive dementia of the central nervous system. The transmissible agent of CJD is unusual for several reasons: It has a long incubation period, up to eight years in some cases; it does not provoke an inflammatory reaction within infected tissues; it has never been isolated; it is resistant to routine paraffin processing. Histotechnologists need to be informed of the potential hazards and of the correct handling of tissues infected with this unconventional agent, even though it has low transmissibility. Treatment of CJD tissues with long sterilization at high temperatures or prolonged exposure to bleach has proven effective in deactivating the agent. This article outlines the precautionary procedures used in our laboratory when processing CJD tissues. (J Histotechnol12:214, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.214

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractA special staining technique for demonstratingHistoplasma duboisiiin paraffin sections was developed during a study on the staining characteristics of this fungus, which acts as the causative organism for a type of fungal granuloma known as African histoplasmosis, found mostly in African countries. The method is relatively simple, reliable, and easily reproducible. It uses two common dyes, eosin and Alcian blue, that are readily available. The technique does not require any special fixatives and can be carried out on formalin fixed, parafin processed specimens. Because the method also eliminates the complicated and time-consuming special techniques previously used to demonstrate this fungus, it is more economical and time-saving. In paraffin sections,H. duboisiiare demonstrated as round to ovoid sky-blue circlets in rose-pink background. (J Histotechnol12:220, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.220

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractA modified alizarin red S and a silver nitrate method for the demonstration of calcium are described. In the alizarin red S method, isopropyl alcohol is used to dehydrate the sections, thus minimizing the solubility of the calcium lake. The sensitive silver method, based on sensitization and physical development, demonstrates minute traces of calcium deposits mainly in the form of phosphates and is comparable to the alizarin red S. (J Histotechnol12:225, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.225

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor

摘要:

AbstractA case of cytomegalovirus (CMV) infection of the bladder is described, with particular reference to the use ofIn Situhybridization as a diagnostic aid. Morphologically, it was not possible to identify characteristic cytomegalic cells in the tissue. Immunocytochemistry using human serum from an infected patient gave equivocal staining due to cross reaction of the labeled anti-human immunoglobulin serum with tissue proteins. A commercially available monoclonal antibody produced clearer results.In Situhybridization with biotin labeled CMV DNA was able to detect virus unambiguously in routinely prepared paraffin embedded tissue sections. The technique is based on the use of a high temperature to denature tissue DNA sequences, followed by simple immunological detection of hybridized biotin residues. We suggest thatIn Situhybridization should be considered as a diagnostically useful tool in the histological investigation of viral infection. (The J Histotechnol12:229, 1989).

ISSN:0147-8885    

DOI:10.1179/his.1989.12.3.229

出版商:Taylor&Francis    

年代:1989

数据来源: Taylor