Fluctuation-Based Super-Resolution Traction Force Microscopy,Nano Letters

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Fluctuation-Based Super-Resolution Traction Force Microscopy
Nano Letters ( IF 9.6 ) Pub Date : 2020-03-06 , DOI: 10.1021/acs.nanolett.9b04083
Aki Stubb ₁ , Romain F Laine _{2,

3} , Mitro Miihkinen ₁ , Hellyeh Hamidi ₁ , Camilo Guzmán ₁ , Ricardo Henriques _{2,

3} , Guillaume Jacquemet _{1,

4} , Johanna Ivaska _{1,

5}

Affiliation

Cellular mechanics play a crucial role in tissue homeostasis and are often misregulated in disease. Traction force microscopy is one of the key methods that has enabled researchers to study fundamental aspects of mechanobiology; however, traction force microscopy is limited by poor resolution. Here, we propose a simplified protocol and imaging strategy that enhances the output of traction force microscopy by increasing i) achievable bead density and ii) the accuracy of bead tracking. Our approach relies on super-resolution microscopy, enabled by fluorescence fluctuation analysis. Our pipeline can be used on spinning-disk confocal or widefield microscopes and is compatible with available analysis software. In addition, we demonstrate that our workflow can be used to gain biologically relevant information and is suitable for fast long-term live measurement of traction forces even in light-sensitive cells. Finally, using fluctuation-based traction force microscopy, we observe that filopodia align to the force field generated by focal adhesions.

中文翻译：

基于波动的超分辨率牵引力显微镜

细胞力学在组织稳态中发挥着至关重要的作用，并且在疾病中经常被错误调节。牵引力显微镜是研究人员研究机械生物学基本方面的关键方法之一；然而，牵引力显微镜受到分辨率差的限制。在这里，我们提出了一种简化的协议和成像策略，通过增加i）可实现的珠子密度和ii）珠子跟踪的准确性来增强牵引力显微镜的输出。我们的方法依赖于通过荧光波动分析实现的超分辨率显微镜。我们的管道可用于转盘共焦或宽视野显微镜，并且与可用的分析软件兼容。此外，我们证明我们的工作流程可用于获取生物学相关信息，并且即使在光敏感细胞中也适用于牵引力的快速长期实时测量。最后，使用基于波动的牵引力显微镜，我们观察到丝状伪足与粘着斑产生的力场对齐。

更新日期：2020-04-24

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