ETH Zurich, Department of Biosystems Science and Engineering, Basel, Switzerland.
Nat Protoc. 2012 Jan 5;7(1):143-54. doi: 10.1038/nprot.2011.434.
To understand the role of physical forces at a cellular level, it is necessary to track mechanical properties during cellular processes. Here we present a protocol that uses flat atomic force microscopy (AFM) cantilevers clamped at constant height, and light microscopy to measure the resistance force, mechanical stress and volume of globular animal cells under compression. We describe the AFM and cantilever setup, live cell culture in the AFM, how to ensure stability of AFM measurements during medium perfusion, integration of optical microscopy to measure parameters such as volume and track intracellular dynamics, and interpretation of the physical parameters measured. Although we use this protocol on trypsinized interphase and mitotic HeLa cells, it can also be applied to other cells with a relatively globular shape, especially animal cells in a low-adhesive environment. After a short setup phase, the protocol can be used to investigate approximately one cell per hour.
为了了解细胞水平上物理力的作用，有必要在细胞过程中跟踪机械性能。在这里，我们提出了一种使用固定高度夹持的平面原子力显微镜（AFM）悬臂来测量球形动物细胞在压缩下的阻力、机械应力和体积的方案。我们描述了 AFM 和悬臂的设置、AFM 中的活细胞培养、如何确保在介质灌注过程中 AFM 测量的稳定性、将光学显微镜集成以测量体积等参数并跟踪细胞内动力学，以及解释所测量的物理参数。尽管我们在胰蛋白酶处理的间期和有丝分裂 HeLa 细胞上使用此方案，但它也可以应用于其他具有相对球形的细胞，特别是在低粘附环境中的动物细胞。在短的设置阶段之后，该方案可以用于每小时大约研究一个细胞。
