Live-cell subcellular measurement of cell stiffness using a microengineered stretchable micropost array membrane.
Live-cell subcellular measurement of cell stiffness using a microengineered stretchable micropost array membrane.
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DOI：
10.1039/c2ib20134h
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发表时间：
2012-10
期刊：
影响因子：
--
通讯作者：
Fu J
中科院分区：
文献类型：
--
作者：
Lam RH;Weng S;Lu W;Fu J
力越来越多地被认为是细胞结构和功能的主要调节因子，并且细胞的机械特性（例如细胞刚度）对于细胞感知力、将力传递到细胞内部或其他细胞并将其转化为影响细胞反应谱的化学信号的机制是必不可少的。在这里，我们报告了一种新的全细胞细胞刚度测量技术与亚细胞空间分辨率。这项技术是基于一种新的细胞拉伸装置，允许定量控制和实时测量的机械刺激和细胞生物力学反应。我们的策略涉及集成到可拉伸弹性膜上的硅橡胶弹性体微柱的微制造阵列。使用计算机控制的真空，该微柱阵列膜（mPAM）被激活以向附着在微柱顶部上的粘附细胞施加等双轴细胞拉伸力。使用荧光显微镜记录mPAM拉伸之前和之后的微柱顶部位置，并进一步用于量化局部细胞拉伸力和细胞面积增量。一个强大的计算方案的开发和实施的亚细胞定量的细胞刚度使用的数据的局部细胞拉伸力和细胞面积增量产生的mPAM细胞拉伸试验。我们的细胞刚度的研究，使用mPAM揭示了细胞刚度，细胞牵引力，细胞铺展面积之间的强正相关性，并说明了重要的功能作用，肌动蛋白聚合和肌球蛋白II介导的细胞骨架收缩调节细胞刚度。总的来说，我们的工作报告了一种具有亚细胞空间分辨率的全细胞细胞刚度测量的新方法，这可能有助于解释细胞的复杂生物力学功能和力传感机制，并为细胞和组织工程以及其他体内应用设计更好的材料。
Forces are increasingly recognized as major regulators of cell structure and function, and the mechanical properties of cells, such as cell stiffness, are essential to the mechanisms by which cells sense forces, transmit them to the cell interior or to other cells, and transduce them into chemical signals that impact a spectrum of cellular responses. Here we reported a new whole-cell cell stiffness measurement technique with a subcellular spatial resolution. This technique was based on a novel cell stretching device that allowed for quantitative control and real-time measurements of mechanical stimuli and cellular biomechanical responses. Our strategy involved a microfabricated array of silicone elastomeric microposts integrated onto a stretchable elastomeric membrane. Using a computer-controlled vacuum, this micropost array membrane (mPAM) was activated to apply equibiaxial cell stretching forces to adherent cells attached on the tops of the microposts. The micropost top positions before and after mPAM stretches were recorded using fluorescence microscopy and further utilized to quantify local cell stretching forces and cell area increments. A robust computation scheme was developed and implemented for subcellular quantifications of cell stiffness using the data of local cell stretching forces and cell area increments generated from mPAM cell stretch assays. Our cell stiffness studies using the mPAM revealed strong positive correlations among cell stiffness, cellular traction force, and cell spread area, and illustrated the important functional roles of actin polymerization and myosin II-mediated cytoskeleton contractility in regulating cell stiffness. Collectively, our work reported a new approach for whole-cell cell stiffness measurements with a subcellular spatial resolution, which would likely help explain the complex biomechanical functions and force-sensing mechanisms of cells and design better materials for cell and tissue engineering and other applications in vivo.
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影响因子：
3.4
作者：
Gavara, Nuria;Roca-Cusachs, Pere;Navajas, Daniel
通讯作者：
Navajas, Daniel
影响因子：
2.4
作者：
Mathur, AB;Collinsworth, AM;Truskey, GA
通讯作者：
Truskey, GA
DOI：
10.1126/science.1191035
发表时间：
2010-08-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gilbert PM;Havenstrite KL;Magnusson KE;Sacco A;Leonardi NA;Kraft P;Nguyen NK;Thrun S;Lutolf MP;Blau HM
通讯作者：
Blau HM
影响因子：
3.7
作者：
Hsu HJ;Lee CF;Locke A;Vanderzyl SQ;Kaunas R
通讯作者：
Kaunas R
影响因子：
6.1
作者：
Mann JM;Lam RH;Weng S;Sun Y;Fu J
通讯作者：
Fu J