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利用钩状原子力显微镜探针直接操纵细胞内的应激纤维。

Direct manipulation of intracellular stress fibres using a hook-shaped AFM probe.

机构信息

Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama, Kanagawa, Japan.

出版信息

Nanotechnology. 2010 Sep 24;21(38):385102. doi: 10.1088/0957-4484/21/38/385102. Epub 2010 Aug 27.

Abstract

Atomic force microscopy (AFM) is a highly successful technique for imaging nanometre-sized samples and measuring pico- to nano-newton forces acting between atoms and molecules. When it comes to the manipulation of larger samples with forces of tens and hundreds of nano-newtons, however, the present chemistry-based modification protocols for functionalizing AFM cantilevers to achieve the formation of covalent/non-covalent linkages between the AFM probe and the sample surface do not produce strong enough bonds. For the purpose of measuring the fracture strength and other mechanical properties of stress fibres (SFs) in living as well as semi-intact fibroblast cells, we fabricated an AFM probe with a hooking function by focused ion beam technology and used the AFM probe hook to capture, pull and eventually sever a chosen SF labelled with green or red fluorescent protein.

摘要

原子力显微镜(AFM)是一种非常成功的技术,可用于成像纳米级大小的样品,并测量原子和分子之间相互作用的皮牛到纳牛的力。然而,当涉及到用数十到数百纳牛顿的力操纵更大的样品时,目前用于对 AFM 悬臂进行功能化的基于化学的修饰方案,无法产生足够强的键来实现 AFM 探针与样品表面之间的共价/非共价键合。为了测量活细胞和半完整成纤维细胞中的应力纤维(SF)的断裂强度和其他机械性能,我们使用聚焦离子束技术制造了具有钩状功能的 AFM 探针,并使用 AFM 探针钩捕获、拉动并最终切断用绿色或红色荧光蛋白标记的选定 SF。

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