Bufi Nathalie, Durand-Smet Pauline, Asnacios Atef
Laboratoire Matières et Systèmes Complexes, Université Paris-Diderot/CNRS, Sorbonne Paris Cité, Paris, France.
Methods Cell Biol. 2015;125:187-209. doi: 10.1016/bs.mcb.2014.11.002. Epub 2015 Jan 9.
We describe here the parallel plates technique which enables quantifying single-cell mechanics, either passive (cell deformability) or active (whole-cell traction forces). Based on the bending of glass microplates of calibrated stiffness, it is easy to implement on any microscope, and benefits from protocols and equipment already used in biology labs (coating of glass slides, pipette pullers, micromanipulators, etc.). We first present the principle of the technique, the design and calibration of the microplates, and various surface coatings corresponding to different cell-substrate interactions. Then we detail the specific cell preparation for the assays, and the different mechanical assays that can be carried out. Finally, we discuss the possible technical simplifications and the specificities of each mechanical protocol, as well as the possibility of extending the use of the parallel plates to investigate the mechanics of cell aggregates or tissues.
我们在此描述一种平行板技术，该技术能够对单细胞力学进行量化，包括被动力学（细胞变形能力）或主动力学（全细胞牵引力）。基于校准刚度的玻璃微板的弯曲，它易于在任何显微镜上实施，并受益于生物学实验室中已使用的方案和设备（玻片涂层、移液器拉制器、显微操作器等）。我们首先介绍该技术的原理、微板的设计和校准，以及对应于不同细胞-基质相互作用的各种表面涂层。然后我们详细说明用于测定的特定细胞制备方法，以及可以进行的不同力学测定。最后，我们讨论可能的技术简化以及每个力学方案的特异性，以及将平行板的使用扩展到研究细胞聚集体或组织力学的可能性。
