High-Resolution, Highly-Integrated Traction Force Microscopy Software.
High-Resolution, Highly-Integrated Traction Force Microscopy Software.
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DOI：
10.1002/cpz1.233
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发表时间：
2021-09
期刊：
影响因子：
--
通讯作者：
Han SJ
中科院分区：
文献类型：
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作者：
Mittal N;Han SJ
准确测量细胞牵引力对于理解细胞和细胞外基质之间的物理相互作用至关重要。牵引力显微镜（TFM）已成为用于此目的最广泛使用的工具。尽管 TFM 在分辨率和准确性方面取得了不断的发展，但在获得用户友好的软件以及为与软件相关的参数和子过程选择正确的值方面仍然存在挑战。在这里，我们提供了基于 MATLAB 的 TFM 软件的分步说明，该软件配备了图像变形量化和力重建的多种方式，并阐明了软件内参数的计算含义。我们概述了如何根据图像的特征和分析目的选择每个过程的最佳子方法和参数值。对于标准工作站中的单幅图像（1280 × 960 像素），该软件的运行时间分别为 20 分钟、4 分钟和 0.05 分钟，其中快速 BEM L1（边界元法 L1 正则化）、快速 BEM L2（L2 正则化）和 FTTC（傅立叶变换牵引细胞术），以及基于粒子跟踪测速的变形跟踪 7 分钟。最后比较了三种力重建方法之间参考桩蛋白-GFP 图像的共定位精度。
Accurate measurement of the cellular traction force is critical for understanding physical interaction between cells and the extracellular matrix. Traction force microscopy (TFM) has become the most widely used tool for such a purpose. While TFM has made an unceasing development in terms of resolution and accuracy, there have been challenges regarding obtaining user-friendly software and choosing the right values for parameters and sub-processes associated with the software. Here, we provide step-by-step instruction of the MATLAB-based TFM software equipped with multiple ways of the image deformation quantification and the force reconstruction, along with clarification on the computational meaning of the parameters within the software. We outline how to choose the optimal sub-methods and values of parameters per each process depending on the characteristics of images and purpose of the analyses. The software’s running time is 20, 4 and 0.05 minutes by Fast BEM L1 (Boundary Element Method L1-regularization), Fast BEM L2 (L2-regularization) and FTTC (Fourier Transform Traction Cytometry), respectively, in addition to 7 minutes of particle-tracking-velocimetry-based deformation tracking, for a single image (1280 × 960 pixel) in a standard workstation. The colocalization accuracies, in reference to a paxillin-GFP image, are compared between the three force reconstruction methods at the end.