Although immunohistochemistry (IHC) is a popular imaging technique, the quantitative analysis of IHC images via computer-aided methods is an emerging field that is gaining more and more importance thanks to the new developments in digital high-throughput scanners. In this paper, we discuss the main steps of IHC and review the techniques for computer-aided chromogenic IHC analysis, including methods to determine the location of interest of the antigens and quantify their activations. Moreover, we discuss the issues arising from the standardization of the immunostaining process, that are generally overlooked by the current literature, and finally provide requirements for reliable computer-aided IHC quantification.
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