Molecular dynamics and forces of a motile cell simultaneously visualized by TIRF and force microscopies
Molecular dynamics and forces of a motile cell simultaneously visualized by TIRF and force microscopies
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DOI：
10.2144/000112752
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发表时间：
2008-05-01
期刊：
影响因子：
2.7
通讯作者：
Yumura, Shigehiko
中科院分区：
文献类型：
--
作者：
Iwadate, Yoshiaki;Yumura, Shigehiko
细胞必须对基质施加牵引力以持续迁移。分子动力学，如肌动蛋白聚合在前面的细胞和肌球蛋白II的积累在后方发挥重要作用所需的迁移力。因此，揭示分子动力学与牵引力之间的关系具有重要意义。牵引力可以从迁移细胞下面的弹性基底的变形来计算。用一对铂催化的有机硅制成了一种透明无色的高折射率（1.40）、低杨氏模量（1.0kPa）的弹性基底。我们使用这个基质开发了一种新的方法，同时记录分子动力学和牵引力下的迁移细胞，其中全内折射荧光（TIRF）和力显微镜相结合。这种新的方法允许检测的时空分布的牵引力所产生的个别filepodia迁移Dictyosteoblastoma细胞。以及这些牵引力和丝状肌球蛋白II的动力学的同时可视化。
Cells must exert traction forces onto the substratum for continuous migration. Molecular dynamics such as actin polymerization at the front of the cell and myosin II accumulation at the rear should play important roles in the exertion of forces required for migration. Therefore, it is important to reveal the relationships between the traction forces and molecular dynamics. Traction forces can be calculated from the deformation of the elastic substratum Under it migrating cell. A transparent and colorless elastic substratum with a high refractive index (1.40) and it low Young'S modulus (1.0 kPa) were made from a pair of platinum-catalyzed Silicones. We used this substratum to develop a new method for simultaneous recording of molecular dynamics and traction forces under a migrating cell in which total internal refractive flourescence (TIRF) and force microscopies were combined. This new method allows the detection of the spatiotemporal distribution of traction forces produced by individual filopodia in migrating Dictyostelium cells. as well as simulataneous visualization of these traction forces and the dynamics of filamentous myosin II.