scholarly journals THE USE OF BASIC POLYSACCHARIDES IN HISTOCHEMISTRY AND CYTOCHEMISTY: III. ALTERED STAINING OF ACIDIC POLYSACCHARIDES IN TISSUE SECTIONS INDUCED BY ASPERGILLUS POLYSACCHARIDE AND DEACETYLATED CHITIN

1962 ◽  
Vol 10 (1) ◽  
pp. 19-23 ◽  
Author(s):  
SIDNEY P. KENT ◽  
E. EDWARD EVANS

The effect of basic polysaccharides on the staining characteristics of acidic polysaccharides in tissue sections of formalin-fixed paraffinembedded material has been studied. Acidic polysaccharides ordinarily do not stain with acidic dyes, but stain well with basic dyes. The complex formed when a basic polysaccharide such as Aspergillus polysaccharide or deacetylated chitin reacts with acidic polysaccharides assumes the staining characteristics of the basic polysaccharide, that is, it stains with acidic dyes. Thus, areas in tissue sections containing acidic polysaccharides can be colored with acidic dyes. Further, the basic polysaccharides were shown to temporarily block staining of acidic polysaccharides by alcian blue and mucicarmine.

1962 ◽  
Vol 10 (1) ◽  
pp. 14-18 ◽  
Author(s):  
SIDNEY P. KENT ◽  
E. EDWARD EVANS

A method for demonstrating acidic polysaccharides in formalin fixed parafin embedded tissue sections using fluorescein labelled deacetylated chitin is described. Epithelial and connective tissue acidic polysaccharides have been studied in a number of organs. The distribution of acidic polysaccharides as seen with fluorescein-labelled deacetylated chitin is similar to results obtained with alcian blue-periodic acid Schiff and fluorescein-labelled Aspergillus polysaccharide. Previously reported fluorescent staining methods for acidic polysaccharides such as iron-hematoxylin-acridine orange and atabrine as well as fluorescein-labelled deacetylated chitin may prove to be useful adjuncts to the light microscopic methods of demonstrating mucins. Other possible uses of deacetylated chitin, a colorless macro-cation, in histochemistry and cytochemistry are noted.


1962 ◽  
Vol 10 (1) ◽  
pp. 8-13 ◽  
Author(s):  
E. EDWARD EVANS ◽  
SIDNEY P. KENT

A procedure was described for demonstrating acidic polysaccharides in formalin-fixed paraffin-embedded tissue sections using fluorescein-labelled Aspergillus polysaccharide. This material is a basic polysaccharide composed of units of galactosamine and N-acetyl galactosamine and is synthesized by the mold Aspergillus parasiticus. The distribution of acidic polysaccharides shown by the fluorescein-labelled Aspergillus polysaccharide method was similar to results with the alcian blue-periodic acid Schiff sequence and to those with fluorescein labelled deacetylated chitin. Both epithelial and connective tissue mucins were demonstrated by the fluorescein-labelled basic polysaccharide. Aspergillus polysaccharide may be considered a colorless polyvalent cation that acts as a carrier for the fluorochrome in this procedure. Since it can react with many types of biological cells as well as with various substances isolated from cells, it may have other important applications in histochemistry and cytochemistry.


2016 ◽  
Vol 54 (11) ◽  
pp. 2798-2803 ◽  
Author(s):  
Elham Salehi ◽  
Mohammad T. Hedayati ◽  
Jan Zoll ◽  
Haleh Rafati ◽  
Maryam Ghasemi ◽  
...  

In a retrospective multicenter study, 102 formalin-fixed paraffin-embedded (FFPE) tissue specimens with histopathology results were tested. Two 4- to 5-μm FFPE tissue sections from each specimen were digested with proteinase K, followed by automated nucleic acid extraction. Multiple real-time quantitative PCR (qPCR) assays targeting the internal transcribed spacer 2 (ITS2) region of ribosomal DNA, using fluorescently labeled primers, was performed to identify clinically important genera and species of Aspergillus , Fusarium , Scedosporium , and the Mucormycetes . The molecular identification was correlated with results from histological examination. One of the main findings of our study was the high sensitivity of the automated DNA extraction method, which was estimated to be 94%. The qPCR procedure that was evaluated identified a range of fungal genera/species, including Aspergillus fumigatus , Aspergillus flavus , Aspergillus terreus , Aspergillus niger , Fusarium oxysporum , Fusarium solani , Scedosporium apiospermum , Rhizopus oryzae , Rhizopus microsporus , Mucor spp., and Syncephalastrum . Fusarium oxysporum and F. solani DNA was amplified from five specimens from patients initially diagnosed by histopathology as having aspergillosis. Aspergillus flavus , S. apiospermum , and Syncephalastrum were detected from histopathological mucormycosis samples. In addition, examination of four samples from patients suspected of having concomitant aspergillosis and mucormycosis infections resulted in the identification of two A. flavus isolates, one Mucor isolate, and only one sample having both R. oryzae and A. flavus . Our results indicate that histopathological features of molds may be easily confused in tissue sections. The qPCR assay used in this study is a reliable tool for the rapid and accurate identification of fungal pathogens to the genus and species levels directly from FFPE tissues.


1996 ◽  
Vol 44 (11) ◽  
pp. 1251-1259 ◽  
Author(s):  
J R Reeves ◽  
J J Going ◽  
G Smith ◽  
T G Cooke ◽  
B W Ozanne ◽  
...  

The relationship between expression of the c-erbB-2 proto-oncogene and the biology of breast cancer has been investigated widely, most studies using immunohistochemistry in formalin-fixed, paraffin-embedded tissues. This technique is at best semiquantitative and there is a high degree of interstudy variability because of its subjective nature and poor methodological standardization. The relationship between the levels of expression and biology can be examined thoroughly only with an accurately quantitative technique. We have developed a radioimmunohistochemical assay to measure p185(erbB-2) in tissue biopsy specimens. The method involves incubating frozen sections with 125I-labeled monoclonal antibody, microautoradiograpy, and grain counting with image analysis. Sections of cell pellets with known c-erbB-2 levels are processed with each batch of samples as internal calibration standards. We have quantified c-erbB-2 expression in 60 breast carcinomas and compared the results with conventional immunohistochemistry. Radioimmunohistochemistry measured receptor levels throughout the range of expression in breast carcinomas, whereas conventional immunohistochemistry detected the protein only in the highest expressing tumors. The quantitative, objective data produced by radioimmunohistochemistry allow a more thorough evaluation of the relationship between c-erbB-2 expression and tumor biology. This technique may have applications in other fields where quantitative data is required and relevant monoclonal antibodies are available.


1995 ◽  
Vol 52 (3) ◽  
pp. 309-316 ◽  
Author(s):  
Yoshito Eizuru ◽  
Yoichi Minamishima ◽  
Tadashi Matsumoto ◽  
Toshinari Hamakado ◽  
Mikio Mizukoshi ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document