4.2 Immunohistochemistry and Immunocytochemistry
Immunochemistry and immunofluorescence are so closely related to immunoassay that a new chapter has been added to include them. The field of proteomics has brought immunoassay and immunofluorescence closer as both are used to locate and quantify proteins of scientific interest in cells, comparing diseased and normal cells for clues about causation and possible treatments, or to evaluate one treatment against another at the cellular level.
Abstract
The principles and practical implementation of immunohistochemistry and immunocytochemistry are explained. The four common specimen formats are described: paraffin embedded, frozen, free floating and cytological. There is a detailed section on fixation, followed by sections on processing tissue blocks to paraffin wax, the use of microtomy and tissue microarrays. Information is also provided on specimen storage and decalcification. The next section, on antigen retrieval, includes full protocols. Controls are also reviewed. There is a detailed section on optimizing an antibody for immunochemical staining and choosing an appropriate label and signal detection technique, including a table of available fluorochromes. In the next section, protocols are provided for staining and counterstaining. Finally, examples of photomicrographs from immunohistochemistry and immunocytochemistry/immunofluorescence are provided.
Contributor
Simon Renshaw is currently the Senior Imaging Scientist at Abcam plc, Cambridge (UK). Simon completed his Biomedical Sciences degree at the University of Bradford (UK) in 1998. He then trained to be a BMS1 in the Department of Histopathology at Addenbrooke’s NHS Trust in Cambridge (UK), where he was first introduced to using IHC and IF techniques for diagnostic purposes. Simon started at Abcam plc in 2001.
Keywords
Immunochemistry, immunohistochemistry, immunocytochemistry, immunostaining, microtomy, tissue section, tissue block, stain, fixation, tissue microarray, paraffin-embedded section, fluorescence, decalcification, antigen retrieval, controls, antibody, enzyme label, horseradish peroxidase, alkaline phosphatase, signal amplification, fluorochromes, counterstain, mounting, photomicrography.