Multiplex Immunohistochemistry for Mapping the Tumor Microenvironment.

Multiplex Immunohistochemistry (mIHC) allows for the simultaneous detection of multiple targets of interest in a single tissue section. Using this method it is therefore possible to evaluate several effectors of a signal transduction pathway at one time, co-localization of markers, or distribution of molecules relative to one another within tissues. This method plays an important role in characterizing molecular signaling and the outcome of signal ablation when assessing mechanisms of drug action. Additionally, this approach can be used to characterize: the tumor microenvironment including vascular architecture and hypoxia, cellular proliferation, cell death as well as drug distribution. Medium to high-throughput images can be obtained via automation of slide imaging and analysis. Here we describe the basic principles of designing and implementing an mIHC protocol to create parameter tumor maps of a variety of specifically labeled markers.