Immunohistochemistry for structural and functional analysis in cardiovascular research

Abstract

Immunohistochemistry is one of the main methods for specific detection and recognition of molecules in cells and tissue and has a long tradition. The principle of the immunohistochemical method is localization and identification of a specific antigen in a cell or a tissue specimen. Immunohistochemical detection has been done for more then 60 years, starting with the detection of bacteria (Coons et al. 1941). The progression in development of antibodies and a series of technical developments have led to an increased sensitivity in immunohistochemical methods and made immunohistochemistry one of the most essential methods in analytical morphology. In cardiovascular research as in other fields, a large increase in the use of immunohistochemistry can be observed in the last 10 years. Because of the improved methods for immunohistochemistry, its employment has also been expanded. While in the past immunohistochemistry was mainly used for the cellular localization of molecules in the tissue, now detection of subcellular localization, colocalization with other molecules and time-dependent alterations in expression and subcellular localization are the foci of interest. Besides these aspects, a further application becomes more and more the focus of interest, the detection of molecules in the activated and/or phosphorylated form. The rapid development in the immunohistochemical field and the expanded and altered use of immunohistochemistry makes it necessary to choose the immunohistochemical method for the individual application critically. Especially the choices of fixation, primary and secondary antibody and visualization method depend upon the underlying question for an optimal result. An improvement in immunohistochemical results can also be elicited by the use of new methods that allow one to increase the sensitivity and specificity of the immunohistochemistry. © Springer-Verlag Berlin Heidelberg 2005.