Alenghat F J, Fabry B, Tsai K Y, Goldmann W H, Ingber D E
Department of Pathology, Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
Biochem Biophys Res Commun. 2000 Oct 14;277(1):93-9. doi: 10.1006/bbrc.2000.3636.
A magnetic tweezer was constructed to apply controlled tensional forces (10 pN to greater than 1 nN) to transmembrane receptors via bound ligand-coated microbeadswhile optically measuring lateral bead displacements within individual cells. Use of this system with wild-type F9 embryonic carcinoma cells and cells from a vinculin knockout mouse F9 Vin (-/-) revealed much larger differences in the stiffness of the transmembrane integrin linkages to the cytoskeleton than previously reported using related techniques that measured average mechanical properties of large cell populations. The mechanical properties measured varied widely among cells, exhibiting an approximately log-normal distribution. The median lateral bead displacement was 2-fold larger in F9 Vin (-/-) cells compared to wild-type cells whereas the arithmetic mean displacement only increased by 37%. We conclude that vinculin serves a greater mechanical role in cells than previously reported and that this magnetic tweezer device may be useful for probing the molecular basis of cell mechanics within single cells.
构建了一种磁性镊子，通过结合配体包被的微珠，对跨膜受体施加可控张力（10皮牛至大于1纳牛），同时光学测量单个细胞内微珠的横向位移。将该系统应用于野生型F9胚胎癌细胞和纽蛋白基因敲除小鼠F9 Vin（-/-）的细胞，结果显示，与之前使用测量大细胞群体平均力学特性的相关技术相比，跨膜整合素与细胞骨架连接的刚度差异要大得多。测量的力学特性在细胞间差异很大，呈现出近似对数正态分布。与野生型细胞相比，F9 Vin（-/-）细胞中微珠的横向位移中位数大2倍，而算术平均位移仅增加37%。我们得出结论，纽蛋白在细胞中的力学作用比之前报道的更大，并且这种磁性镊子装置可能有助于探究单个细胞内细胞力学的分子基础。
