Measurement of Minute Cellular Forces by Traction Force Microscopy.

The ability of living cells to exert forces on their surrounding environment, such as the extracellular matrix (ECM) or neighboring cells, plays an important role in numerous biological processes. This chapter describes a simple protocol to measure forces exerted by living cells using traction force microscopy. This approach is based on the measurement of the deformation of compliant substrates using fluorescent fiducials. It can be implemented using widefield or confocal fluorescent microscopes, and open-source software. This chapter describes a step-by-step protocol to measure forces exerted by focal adhesions bound to the ECM protein fibronectin. However, this framework is versatile and can be easily adapted to a multitude of ligands and cellular processes in which cells exert forces, including the formation of an immunological synapse or a phagocytic cup. Technical considerations, limitations of the approach, and practical advice are discussed.