Shear force at the cell-matrix interface: enhanced analysis for microfabricated post array detectors.

Christopher A. Lemmon, N. Sniadecki, Sami Alom Ruiz, J. Tan, L. Romer, Christopher S. Chen
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引用次数: 104

Abstract

The interplay of mechanical forces between the extracellular environment and the cytoskeleton drives development, repair, and senescence in many tissues. Quantitative definition of these forces is a vital step in understanding cellular mechanosensing. Microfabricated post array detectors (mPADs) provide direct measurements of cell-generated forces during cell adhesion to extracellular matrix. A new approach to mPAD post labeling, volumetric imaging, and an analysis of post bending mechanics determined that cells apply shear forces and not point moments at the matrix interface. In addition, these forces could be accurately resolved from post deflections by using images of post tops and bases. Image analysis tools were then developed to increase the precision and throughput of post centroid location. These studies resulted in an improved method of force measurement with broad applicability and concise execution using a fully automated force analysis system. The new method measures cell-generated forces with less than 5% error and less than 90 seconds of computational time. Using this approach, we demonstrated direct and distinct relationships between cellular traction force and spread cell surface area for fibroblasts, endothelial cells, epithelial cells and smooth muscle cells.
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细胞-基质界面的剪切力:微制造柱阵列探测器的增强分析。
细胞外环境和细胞骨架之间的机械力的相互作用驱动着许多组织的发育、修复和衰老。这些力的定量定义是理解细胞力学传感的重要一步。微加工后阵列探测器(mPADs)提供了细胞产生的力在细胞粘附到细胞外基质的直接测量。一种新的mPAD后标记、体积成像和后弯曲力学分析方法确定了细胞在基质界面施加剪切力而不是点矩。此外,利用柱顶和柱底的图像可以准确地从柱的偏转中分辨出这些力。然后开发了图像分析工具,以提高后质心定位的精度和吞吐量。这些研究产生了一种改进的力测量方法,使用全自动力分析系统,具有广泛的适用性和简洁的执行。新方法测量细胞产生的力,误差小于5%,计算时间小于90秒。利用这种方法,我们证明了成纤维细胞、内皮细胞、上皮细胞和平滑肌细胞的细胞牵引力与扩散细胞表面积之间存在直接而独特的关系。
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