Using Antibodies in Microscopy: A Guide to Immunohistochemistry. Part 1: Antibodies, Tissue Fixation, Embedding, and Sectioning. Issue 2 (12th April 2023) - Guerin, Christopher J
Abstract: Immunohistochemistry (IHC) is the use of antibodies as probes to determine the localization of proteins in a cell or tissue sample. In microscopy, the technique has been in use since the 1940s by pathologists and research scientists. At first glance it appears to be a straightforward procedure. In practice, however, an IHC protocol has several steps where problems can occur, resulting in either false positive or false negative data. IHC can be used in both transmitted light and fluorescence microscopy, and it can be used in combinations to identify and mark multiple proteins of interest. While many IHC techniques have been published, a certain amount of confusion exists as to how to optimize IHC procedures. This series of three articles will describe the steps involved in IHC, the function of each step, and some variations that occur in specific protocols on differing cell and tissue types. It is hoped that a better understanding of these steps will help to guide users in determining the optimal conditions for carrying through IHC protocols.