The Post-embedding Method for Immunoelectron Microscopy of Mammalian Tissues: A Standardized Procedure Based on Heat-Induced Antigen Retrieval

Author(s):  
Shuji Yamashita
1998 ◽  
Vol 4 (S2) ◽  
pp. 1114-1115
Author(s):  
John P. Petrali ◽  
Kenneth R. Mills

Microwave energy (MWE) as a method of rapid tissue processing is gaining increasing support as an alternative to routine chemical processing in diagnostic laboratory environments. Chemical fixatives and conventional fixation times used for standardized preservation of tissues can result in serious alterations in morphology as a consequence of solubilization and conformational changes of proteins and lipids. These untoward changes typically result in compromised antigenicity of many tissue proteins. With MWE processing, tissue antigens can be distinctly better preserved, antigen retrieval made more replicate and histochemical and immunochemical reactions made ultrafast. The present study had two objectives: to compare MWE processed skin to conventionally processed skin for ultrastructural integrity; to compare MWE expression of antigenicity of selected skin proteins which are usually sensitive to conventional chemical processing. Skin proteins selected were bullous pemphigoid antigen (BPA) and laminin.


Author(s):  
D. J. McComb ◽  
J. Beri ◽  
F. Zak ◽  
K. Kovacs

Gonadotroph cell adenomas of the pituitary are infrequent in human patients and are not invariably associated with altered gonadal function. To date, no animal model of this tumor type exists. Herein, we describe spontaneous gonadotroph cell adenomas in old male and female Sprague-Dawley rats by histology, immunocytology and electron microscopy.The material consisted of the pituitaries of 27 male and 38 female Sprague Dawley rats, all 26 months of age or older, removed at routine autopsy. Sections of formal in-fixed, paraffin-embedded tissue were stained with hematoxylin-phloxine-saffron (HPS), the PAS method and the Gordon-Sweet technique for the demonstration of reticulin fibers. For immunostaining, sections were exposed to anti-rat β-LH, anti-ratβ-TSH, anti-rat PRL, anti-rat GH and anti-rat ACTH 1-39. For electron microscopy, tissue was fixed in 2.5% glutaraldehyde, postfixed in 1% OsO4 and embedded in epoxy-resin. Tissue fixed in 10% formalin, embedded in epoxy resin without osmification, was used for immunoelectron microscopy.


Author(s):  
K. Pegg-Feige ◽  
F. W. Doane

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.


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