Live Cell Imaging and Analysis of Cancer-Cell Transmigration Through Endothelial Monolayers

During metastasis, a subset of cancer cells will break away from the primary tumor and invade into the surrounding tissue. Cancer cells which are able to breach the endothelium and enter the blood stream are then transported in the circulation to new target organs where they may seed as a distant metastasis. In order to invade this new organ, the cancer cells must bind to and traverse the vascular wall, a process known as transendothelial migration (TEM) or extravasation. This chapter describes an in vitro approach to automated live cell imaging and analysis of TEM in order to accurately quantify these kinetics and aid the researcher in dissecting the mechanisms of tumor-endothelial interactions during this phase of metastasis.