We describe three different methods of calibration of quantitative immunohistochemistry, two of them based on self-made dye-agarose or antigen-agarose substrates. Quantification of immunostained objects (immunosignals) can be done by (1) microscope photometry (plug method) or scanning-photometry; (2) and densitometry of a television transformed microscope image. Both approaches are subjected to the different methods of calibration and three examples of application.