ISSN: 2041-6792

Clinical Investigation

Scholarly Journals In Immunhistochemistry

Immunohistochemistry (IHC) is a method to identify specific antigens within tissue sections utilizing an antigen-specific antibody. Detection at the light microscopic level of antigen–antibody interactions can be achieved by labeling the antibody with a substance that can be visualized, either by conjugation to a fluorescent marker or enzyme followed by colorimetric detection. Immunologic detection of antigens dates to the early 20th century when Marrack demonstrated that anti-typhoid and anti-cholera sera-labeled with diazotized benzidine-azo-r-salt imparted a red color to the bacteria. Although groundbreaking for immunological detection of antigens, Coons determined this labeling method to be relatively insensitive when applied to tissues, and subsequently described assays utilizing fluorescent-labeled antibodies in fixed tissues, but interpretation was confounded by the enhanced endogenous fluorescent activity in formalin-fixed tissue. In 1966, Nakane described a method of antigen detection in tissue using an antibody conjugated to an enzyme (horseradish peroxidase) and utilized a colorimetric substrate that could be detected by light microscopy, which is the theoretical basis of most modern tissue-based immunohistochemical assays. This chapter will focus on detection of rabies virus (RABV) antigens in formalin-fixed, paraffin-embedded (FFPE) tissues. The materials and methods describe a single protocol.  However, the reader is encouraged to investigate the many alternative non-proprietary and proprietary protocols that are also available. Detection of antigens in FFPE tissues presents a unique diagnostic challenge regarding validation of the assay and interpretation of the results.    

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