A Biologist’s Guide to Traction Force Microscopy Using Polydimethylsiloxane Substrate for Two-Dimensional Cell Cultures

Summary Cellular traction forces influence epithelial behavior, including wound healing and cell extrusion. Here, we describe a simple in vitro traction force microscopy (TFM) protocol using ECM protein-coated polydimethylsiloxane substrate and widefield fluorescence microscopy. We include detailed steps for analysis so readers can obtain traction forces to study the mechanobiology of epithelial cells. We also provide guidelines on when to adopt another common class of TFM protocols based on polyacrylamide hydrogels. For complete details on the use and execution of this protocol, please refer to Saw et al. (2017) and Teo et al. (2020) .