Introduction to Immunohistochemistry




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It is often useful to be able to stain for two or more antigens in one common tissue section. This can be achieved by immunofluorescence method using different fluorescent dyes. Multiple staining can also be done with peroxidase conjugated antibodies developed with different chromogen substrates to produce the end products of different colors. There are three basic approaches in planning multiple staining: parallel, sequential and adjacent. In addition, the antibody dilution and condition are also important factors to be considered.  Finally, appropriate color combination is also crucial since improper color combination may produce poor result and fail to demonstrate multiple antigens in the same section. For best result, the careful design and test of multiple staining protocols are necessary.