Department of Molecular biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada.
Methods Mol Biol. 2022;2440:125-139. doi: 10.1007/978-1-0716-2051-9_8.
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.
活细胞对其周围环境（如细胞外基质 (ECM) 或相邻细胞）施加力的能力在许多生物过程中起着重要作用。本章描述了一种使用牵引力显微镜测量活细胞施加力的简单方案。该方法基于使用荧光基准标记测量顺应性基底的变形。它可以使用宽场或共聚焦荧光显微镜以及开源软件来实现。本章描述了一种逐步方案，用于测量与 ECM 蛋白纤维连接蛋白结合的粘着斑施加的力。然而，该框架具有通用性，并且可以轻松适应多种配体和细胞施加力的过程，包括免疫突触或吞噬杯的形成。讨论了技术考虑因素、该方法的局限性和实际建议。
