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Nanonet Force Microscopy for Measuring Cell Forces

机译:用于测量细胞力的纳米纳米力显微镜

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摘要

The influence of physical forces exerted by or felt by cells on cell shape, migration, and cytoskeleton arrangement is now widely acknowledged and hypothesized to occur due to modulation of cellular inside-out forces in response to changes in the external fibrous environment (outside-in). Our previous work using the non-electrospinning Spinneret-based Tunable Engineered Parameters' suspended fibers has revealed that cells are able to sense and respond to changes in fiber curvature and structural stiffness as evidenced by alterations to focal adhesion cluster lengths. Here, we present the development and application of a suspended nanonet platform for measuring C2C12 mouse myoblast forces attached to fibers of three diameters (250, 400, and 800 nm) representing a wide range of structural stiffness (3-50 nN/mu m). The nanonet force microscopy platform measures cell adhesion forces in response to symmetric and asymmetric external perturbation in single and cyclic modes. We find that contractility-based, inside-out forces are evenly distributed at the edges of the cell, and that forces are dependent on fiber structural stiffness. Additionally, external perturbation in symmetric and asymmetric modes biases cell-fiber failure location without affecting the outside-in forces of cell-fiber adhesion. We then extend the platform to measure forces of (1) cell-cell junctions, (2) single cells undergoing cyclic perturbation in the presence of drugs, and (3) cancerous single-cells transitioning from a blebbing to a pseudopodial morphology.
机译:在细胞形状,迁移和细胞骨架装置上施加或毡被施加的物理力或感觉到的物理力的影响现在被广泛地确认和假设发生,因为响应于外纤维环境的变化(外部)的细胞内输出力的调节而发生)。我们以前的工作采用非静电纺丝喷丝网的可调工程参数'悬浮纤维已透露,细胞能够感测和响应纤维曲率和结构刚度的变化,如通过改变对局部粘合簇长度的改变所证明的。在这里,我们介绍了悬浮纳米型平台的开发和应用,用于测量附着在三径(250,400和800nm)的纤维上的C2C12小鼠肌细胞力,其代表各种结构刚度(3-50 nn / mu m) 。纳米键力显微镜平台响应单循环模式的对称和不对称外部扰动来测量细胞粘附力。我们发现基于收缩的内外力均匀地分布在细胞的边缘,并且该力取决于纤维结构刚度。另外,对称和不对称模式的外部扰动偏置细胞纤维失效位置,而不影响细胞纤维粘附的外部力。然后,我们延长平台以测量(1)细胞 - 细胞结的力,(2)在药物存在下进行循环扰动的单细胞,(3)从BLEBBIBE转换到假缺乏形态的癌细胞单细胞。

著录项

  • 来源
    《Biophysical Journal》 |2016年第1期|共11页
  • 作者单位

    Virginia Tech Dept Biomed Engn &

    Sci Blacksburg VA 24061 USA;

    Virginia Tech Dept Biomed Engn &

    Sci Blacksburg VA 24061 USA;

    Virginia Tech Dept Aerosp &

    Ocean Engn Blacksburg VA 24061 USA;

    Virginia Tech Dept Aerosp &

    Ocean Engn Blacksburg VA 24061 USA;

    Virginia Tech Dept Biomed Engn &

    Sci Blacksburg VA 24061 USA;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物物理学;
  • 关键词

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