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首页> 外文期刊>Acta biomaterialia >A micropatterning and image processing approach to simplify measurement of cellular traction forces
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A micropatterning and image processing approach to simplify measurement of cellular traction forces

机译：一种微图案和图像处理方法，可简化细胞牵引力的测量

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摘要

Quantification of the traction forces that cells apply to their surroundings has been critical to the advancement of our understanding of cancer, development and basic cell biology. This field was made possible through the development of engineered cell culture systems that permit optical measurement of cell-mediated displacements and computational algorithms that allow conversion of these displacements into stresses and forces. Here, we present a novel advancement of traction force microscopy on polyacrylamide (PAA) gels that addresses limitations of existing technologies. Through an indirect patterning technique, we generated PAA gels with fluorescent 1 μm dot markers in a regularized array. This improves existing traction measurements since (i) multiple fields of view can be measured in one experiment without the need for cell removal; (ii) traction vectors are modeled as discrete point forces, and not as a continuous field, using an extremely simple computational algorithm that we have made available online; and (iii) the pattern transfer technique is amenable to any of the published techniques for producing patterns on glass. In the future, this technique will be used for measuring traction forces on complex patterns with multiple, spatially distinct ligands in systems for applying strain to the substrate, and in sandwich cultures that generate quasi-three-dimensional environments for cells.
机译：量化细胞对周围环境施加的牵引力对于提高我们对癌症，发育和基本细胞生物学的理解至关重要。通过开发可对细胞介导的位移进行光学测量的工程化细胞培养系统以及将这些位移转换为应力和力的计算算法，该领域成为可能。在这里，我们介绍了聚丙烯酰胺（PAA）凝胶上的牵引力显微镜技术的新进展，解决了现有技术的局限性。通过间接构图技术，我们在规则阵列中生成了带有荧光1μm点标记的PAA凝胶。由于（i）可以在一个实验中测量多个视野而无需去除细胞，因此可以改善现有的牵引力测量; （ii）使用我们在线提供的极其简单的计算算法，将牵引矢量建模为离散的点力，而不是连续的场; （iii）图案转印技术适合于任何已公开的在玻璃上产生图案的技术。将来，该技术将用于在将应变施加至底物的系统中以及在为细胞生成准三维环境的三明治培养物中，测量具有多个空间上不同的配体的复杂图案上的牵引力。

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