Development of a simple embedding procedure allowing immunocytochemical localization at the ultrastructural level.
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.
Escolar
G
G
Department of Laboratory Medicine, University of Minnesota Health Sciences Center, Minneapolis 55455.
Sauk
J J
JJ
Bravo
M L
ML
Krumwiede
M
M
White
J G
JG
D000179:Acrylates; D001792:Blood Platelets / Q000648:ultrastructure*; D003102:Colloids; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D020366:Methylmethacrylate; D008768:Methylmethacrylates; D008870:Microtubules / Q000648:ultrastructure; D014404:Tubulin / Q000032:analysis
D000179:Acrylates; D003102:Colloids; D008768:Methylmethacrylates; D014404:Tubulin; D020366:Methylmethacrylate