KENBIKYO Vol.49▶No.2 2014
â– Lectures

Immunoelectron Microscopy with the Post-Embedding Method Based on Heat-Induced Antigen Retrieval: Techniques and Mechanisms

Shuji Yamashita

Abstract: Heat-induced antigen retrieval (HIAR) methods that denature protein structure and expose epitopes in fixed tissues are commonly used for immunohistochemistry in recent years, whereas preservation of protein structure has been thought to be important. In this paper, I discussed the mechanisms of HIAR in the tissues fixed with formaldehyde or with glutaraldehyde and osmium tetroxide and described the application of HIAR to immunoelectron microscopy using specimens embedded in LR-White and epoxy resins. In addition, I demonstrated that the selection of diluents for the primary and secondary antibodies is one of the important factors to obtain a sensitive immunoreaction with low background staining.

Key words: Heat-induced antigen retrieval, Immunoelectron microscopy, LR-white resin, Epoxy resin, Diluents for antibodies