Multiple Immunochemical Staining Techniques

Immunochemical staining of cells and tissues is a powerful technique that has emerged in cell biology to provide a direct visualization of proteins and a better understanding of the basic cellular functions. This chapter describes the most common methods employed to successfully perform multiple immunochemical staining using brightfield, conventional epi-fluorescence and confocal microscopy. Multiple immunochemical staining allows the detection of two or more antigens within the same cell or tissue section, thus giving important simultaneous information on the cellular/sub-cellular location, biological function and relationships of various proteins. The chapter focuses on how to choose the optimal staining method, the combination of antibodies and the visualization of the target. These are important aspects to consider when designing a multiple immunochemical staining experiment. Antigen retrieval is often essential for the demonstration of antigens. Multiple reporter label immunochemical staining techniques are considered as advanced immunochemical staining techniques.