Post-embedding Mammalian Tissue for Immunoelectron Microscopy: A Standardized Procedure Based on Heat-Induced Antigen Retrieval

2016 ◽  
pp. 279-290 ◽  
Author(s):  
Shuji Yamashita
Keyword(s):  
Immunoelectron Microscopy ◽  
Antigen Retrieval ◽  
Mammalian Tissue

Microwave-Assisted Immunoelectron Microscopy of Skin

1998 ◽  
Vol 4 (S2) ◽  
pp. 1114-1115
Author(s):  
John P. Petrali ◽  
Kenneth R. Mills
Keyword(s):  
Bullous Pemphigoid ◽  
Immunoelectron Microscopy ◽  
Conformational Changes ◽  
Microwave Energy ◽  
Antigen Retrieval ◽  
Chemical Processing ◽  
Diagnostic Laboratory ◽  
Microwave Assisted ◽  
Bullous Pemphigoid Antigen ◽  
Tissue Antigens

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.

Ultrastructural investigation of spontaneous pituitary gonadotroph cell adenomas in aging Sprague-Dawley rats

1983 ◽  
Vol 41 ◽  
pp. 702-703
Author(s):  
D. J. McComb ◽  
J. Beri ◽  
F. Zak ◽  
K. Kovacs
Keyword(s):  
Electron Microscopy ◽  
Animal Model ◽  
Epoxy Resin ◽  
Immunoelectron Microscopy ◽  
Tumor Type ◽  
Gonadal Function ◽  
Sprague Dawley ◽  
Male And Female ◽  
Reticulin Fibers ◽  
Sprague Dawley Rats

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.

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

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.

Detection of Viral Antigens in Mouse and Rat Tumor Cells by Immunoelectron Microscopy Following Periodate-Lysine-Paraformaldehyde (PLP) Fixation

1976 ◽  
Vol 34 ◽  
pp. 252-253
Author(s):  
Y. Ohtsuki ◽  
G. Seman ◽  
J. M. Bowen ◽  
M. Scanlon ◽  
L. Dmochowski
Keyword(s):  
Immunoelectron Microscopy ◽  
Mammary Tumor ◽  
Type A ◽  
Rabbit Serum ◽  
Virus Particles ◽  
Viral Antigens ◽  
Culture Cells ◽  
Mouse Mammary Tumor ◽  
Tumor Virus ◽  
Immunoperoxidase Labeling

Recently, periodate-lysine-paraformaldehyde (PLP) fixation was reported for immunoelectron microscopy (1). In PLP fixation, carbohydrates are oxidized by periodate and cross-linked by lysine; paraformaldehyde stabilizes proteins and lipids. By using PLP fixation, intracytoplasmic type A viral antigens have been previously demonstrated by immunoperoxidase labeling (2). In the present study, PLP fixation has been applied for the detection of the same antigens in mouse mammary tumor culture cells by both immunoferritin and immunoperoxidase methods. Rabbit anti-intracytoplasmic type A virus serum (anti-A), kindly provided by Dr. M. Muller (3), rabbit anti-strain A mouse mammary tumor virus (anti-MMTV) and preimmune rabbit serum as control were used to detect viral antigens in cells of C3H/HeJ strain mouse mammary tumor culture. Attempts have been also made to demonstrate peroxidase labeling of type C virus particles in frozen sections of an SD-MSV-induced NZB rat bone tumor tissue by rabbit anti-MuLV serum.

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