Schillers Hermann
Institute of Physiology II, University of Münster, Münster, Germany.
Methods Mol Biol. 2019;1886:291-313. doi: 10.1007/978-1-4939-8894-5_17.
Cell's elasticity is an integrative parameter summarizing the biophysical outcome of many known and unknown cellular processes. This includes intracellular signaling, cytoskeletal activity, changes of cell volume and morphology, and many others. Not only intracellular processes defines a cell's elasticity but also environmental factors like their biochemical and biophysical surrounding. Therefore, cell mechanics represents a comprehensive variable of life. A cell in its standard conditions shows variabilities of biochemical and biophysical processes resulting in a certain range of cell's elasticity. Changes of the standard conditions, endogenously or exogenously induced, are frequently paralleled by changes of cell elasticity. Therefore cell elasticity could serve as parameter to characterize different states of a cell. Atomic force microscopy (AFM) combines high spatial resolution with very high force sensitivity and allows investigating mechanical properties of living cells under physiological conditions. However, elastic moduli reported in the literature showed a large variability, sometimes by an order of magnitude (or even more) for the same cell type assessed in different labs. Clearly, a prerequisite for the use of cell elasticity to describe the actual cell status is a standardized procedure that allows obtaining comparable values of a cell independent from the instrument, from the lab and operator. Biologically derived variations of elasticity could not be reduced due to the nature of living cells but technically and methodologically derived variations could be minimized by a standardized procedure.This chapter provides a Standardized Nanomechanical AFM Procedure (SNAP) that reduces strongly the variability of results obtained on soft samples and living cells by a reliable method to calibrate AFM cantilevers.
细胞弹性是一个综合参数,它概括了许多已知和未知细胞过程的生物物理结果。这包括细胞内信号传导、细胞骨架活动、细胞体积和形态的变化等等。不仅细胞内过程决定了细胞的弹性,环境因素如生化和生物物理环境也起着作用。因此,细胞力学是生命的一个综合变量。处于标准条件下的细胞表现出生化和生物物理过程的变异性,导致细胞弹性在一定范围内变化。内源性或外源性诱导的标准条件变化,常常伴随着细胞弹性的变化。因此,细胞弹性可作为表征细胞不同状态的参数。原子力显微镜(AFM)将高空间分辨率与非常高的力灵敏度相结合,能够在生理条件下研究活细胞的力学性质。然而,文献中报道的弹性模量显示出很大的变异性,对于在不同实验室评估的相同细胞类型,有时相差一个数量级(甚至更多)。显然,使用细胞弹性来描述实际细胞状态的一个先决条件是要有一个标准化程序,该程序能够获得与仪器、实验室和操作人员无关的可比细胞值。由于活细胞的性质,生物学衍生的弹性变化无法减少,但通过标准化程序可以将技术和方法学衍生的变化降至最低。本章提供了一种标准化的纳米力学AFM程序(SNAP),通过一种可靠的方法校准AFM悬臂,极大地降低了在软样品和活细胞上获得的结果的变异性。