A Novel Method for Measuring Tension Generated in Stress Fibers by Applying External Forces
A Novel Method for Measuring Tension Generated in Stress Fibers by Applying External Forces
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DOI：
10.1016/j.bpj.2011.05.046
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发表时间：
2011-07-06
影响因子：
3.4
通讯作者：
Sato, Masaaki
中科院分区：
文献类型：
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作者：
Sugita, Shukei;Adachi, Taiji;Sato, Masaaki
细胞骨架应力纤维（SF）中产生的收缩力的分布有助于细胞的动态功能，如迁移和机械转导。这里我们描述一部小说（据我们所知）测量SF中局部张力的方法基于以下步骤：1）在垂直于SF纵轴的方向上对SF施加不同大小的已知力：2）使用力平衡方程来计算SF中由SF角度的变化产生的张力;以及3），将由此建立的张力和所施加的力之间的关系外推到零的所施加的力，以确定SF中预先存在的张力。在这项研究中，我们通过将磁性粒子附着在SF上并用电磁针施加已知的力来测量SF中的张力。荧光显微镜被用来捕捉图像的SF荧光标记的肌球蛋白II抗体，这些图像的分析允许在SF的张力进行测量。在这项研究中测量的平均张力与以前的报告，这表明，这种方法可能成为一个强大的工具，阐明细胞骨架张力影响细胞功能的机制。
The distribution of contractile forces generated in cytoskeletal stress fibers (SFs) contributes to cellular dynamic functions such as migration and mechanotransduction. Here we describe a novel (to our knowledge) method for measuring local tensions in SFs based on the following procedure: 1), known forces of different magnitudes are applied to an SF in the direction perpendicular to its longitudinal axis; 2), force balance equations are used to calculate the resulting tensions in the SF from changes in the SF angle; and 3), the relationship between tension and applied force thus established is extrapolated to an applied force of zero to determine the preexisting tension in the SF. In this study, we measured tensions in SFs by attaching magnetic particles to them and applying known forces with an electromagnetic needle. Fluorescence microscopy was used to capture images of SFs fluorescently labeled with myosin II antibodies, and analysis of these images allowed the tension in the SFs to be measured. The average tension measured in this study was comparable to previous reports, which indicates that this method may become a powerful tool for elucidating the mechanisms by which cytoskeletal tensions affect cellular functions.