Department of Biology, Brooklyn College of the City University of New York, Brooklyn, NY, USA.
Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Methods Mol Biol. 2022;2364:217-235. doi: 10.1007/978-1-0716-1661-1_11.
The aim of this chapter is to present an innovative technique to visualize changes of the F-actin cytoskeleton in response to locally applied force. We developed an in vitro system that combines micromanipulation of force by magnetic tweezers with simultaneous live cell fluorescence microscopy. We applied pulling forces to magnetic beads coated with the Neisseria gonorrhoeae Type IV pili in the same order of magnitude than the forces generated by live bacteria. We saw quick and robust F-actin accumulation in individual cells at the sites where pulling forces were applied. Using the magnetic tweezers, we were able to mimic the local response of the F-actin cytoskeleton to bacteria-generated forces. In this chapter, we describe our magnetic tweezers system and show how to control it in order to study cellular responses to force.
本章旨在介绍一种创新技术，用于可视化 F-肌动蛋白细胞骨架对局部施加力的响应变化。我们开发了一种体外系统，该系统将磁镊对力的微操作与实时细胞荧光显微镜结合在一起。我们以与活细菌产生的力相同的数量级向涂有淋病奈瑟菌 IV 型菌毛的磁性珠施加拉力。我们看到在施加拉力的部位，单个细胞中 F-肌动蛋白迅速而强烈地积累。我们使用磁镊能够模拟 F-肌动蛋白细胞骨架对细菌产生的力的局部响应。在本章中，我们描述了我们的磁镊系统，并展示了如何控制它以研究细胞对力的响应。
