Abstract
The cell mechanical features are largely regulated by actin cytokeleton. By analyzing the mechanical features, it is possible to evaluate the characteristics of the complicated actin cytoskeleton in diverse cell types. In this study, we examined the sub-membrane mechanical structures of normal fibroblasts TIG-1 cells, and cervical cancer Hela cells using local elasticity mapping method of atomic force microscope. Especially we aimed at clarifying the regulatory mechanisms of sub-membrane actin structures in these cells by activation of actomyosin formation using calyculin A. This technique revealed that TIG-1 and Hela cells bore clearly different sub-membrane mechanical structures. TIG-1 cells had aligned stiff filamentous structures, whereas Hela cells had crooked and relatively soft filaments. The surface stiffness of TIG-1 cells increased slightly by actomyosin formation due to stiffness increase of the aligned filamentous structures. On the other hand, the surface stiffness of Hela cells increased by actomyosin formation due to upregulation of the apical actin filaments. Therefore, the structural and regulatory differences of the apical actin filaments could be demonstrated by atomic force microscopy elasticity mapping analysis.
Highlights
The mechanical features of cells are unique indicators for cellular states and diseases
We examined the sub-membrane mechanical structures of normal fibroblasts TIG-1 cells, and cervical cancer Hela cells using local elasticity mapping method of atomic force microscope
It appeared that apical stress fibers of TIG-1 cells decreased a bit, calyculin A treatment did not cause any alterations in the actin cytoskeleton of these cells (Figure 2)
Summary
The mechanical features of cells are unique indicators for cellular states and diseases. There are several methods to measure the mechanical features of cells including micropipette aspiration [5], optical stretcher [6], and atomic force microscopy (AFM) [7]. These technologies measure the surface tension, stiffness, or deformation of cells wholly or locally in physiological conditions. The micropipette aspiration elucidates cell surface tension partially. The optical stretcher elucidates whole cell deformation and AFM measures the cell stiffness or viscoelasticity locally. The spatial resolution of AFM measurements for cell mechanical features is outstanding compared to other methods [8,9,10]. AFM can investigate the mechanical properties of cell surface with high-sensitivity (~1 pN)
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