Using Digital Quantification of Stained Tissue Microarrays as a Medium-Throughput, Quantitative Method for Measuring the Kinetics of Signal Transduction.

Determining how a molecular targeting agent affects signaling pathways is challenging. To facilitate this exploration, we use a combination of medium-throughput methodologies such as tissue microarrays (TMA) and digital image analysis for quantification of immunohistochemistry (IHC). Because TMAs enable the simultaneous analysis of up to 1000 tissues, this tool can be used to comprehensively assess changes in signal transduction pathways over time. TMAs can be digitized and analyzed using the Aperio ScanScope imaging system and Aperio ImageScope software to deliver semiquantitative data. This chapter describes the methodologies for tissue collection and for the construction, sectioning, staining, and digital analysis of TMAs.