Development of a simple embedding procedure allowing immunocytochemical localization at the ultrastructural level.
Author(s) -
Ginés Escolar,
John J. Sauk,
María Luisa Rodes Bravo,
Marcy Krumwiede,
James G. White
Publication year - 1988
Publication title -
journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 124
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1177/36.12.3192938
Subject(s) - polymerization , antigenicity , ultrastructure , methacrylate , benzoyl peroxide , materials science , methyl methacrylate , platelet , acrylic resin , microscopy , polymer chemistry , chemistry , biomedical engineering , nanotechnology , composite material , antigen , biology , anatomy , optics , physics , immunology , polymer , coating , medicine
Immunobed solution A is a water-soluble acrylic compound recently developed for immunocytochemical localization at the light microscopic level. In this study, we combined it with methyl methacrylate (MMA) to achieve sufficient hardness to obtain ultra-thin sections. Samples of platelets were dehydrated and embedded in the water-soluble acrylic mixture (WSAM). The embedding process was carried out at 4 degrees C and final polymerization was induced with either chemical (benzoyl peroxide) or physical (UV light) catalysts. Tubulin was localized at the ultrastructural level in sections embedded according to these two methods. Results were compared with those obtained in platelets processed in Lowicryl. Dehydration and embedding with the WSAM yielded a preservation of antigenicity similar to that obtained in Lowicryl. The new procedure benefits from the low temperature achieved during polymerization, providing good ultrastructural morphology and immunolocalization of protein antigens with the simplicity of a routine embedding procedure for light microscopy.
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