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脂质质膜动力学的超分辨率光学显微镜观察

Super-resolution optical microscopy of lipid plasma membrane dynamics.

机构信息

MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Headley Way, Oxford OX3 9DS, U.K.

出版信息

Essays Biochem. 2015;57:69-80. doi: 10.1042/bse0570069.

Abstract

Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence correlation spectroscopy) on an optical (confocal) microscope is a popular tool for investigating such dynamics. Unfortunately, its full applicability is constrained by the limited spatial resolution of a conventional optical microscope. The present chapter depicts the combination of optical super-resolution STED (stimulated emission depletion) microscopy with FCS, and why it is an important tool for investigating molecular membrane dynamics in living cells. Compared with conventional FCS, the STED-FCS approach demonstrates an improved possibility to distinguish free from anomalous molecular diffusion, and thus to give new insights into lipid-protein interactions and the traditional lipid 'raft' theory.

摘要

质膜动力学是细胞活动的重要调节者,特别是通过膜嵌入蛋白和脂质的相互作用和扩散动力学。在光学(共焦)显微镜上进行的 FCS(荧光相关光谱)是研究此类动力学的常用工具。不幸的是,其完全适用性受到传统光学显微镜空间分辨率的限制。本章描述了光学超分辨率 STED(受激辐射损耗)显微镜与 FCS 的结合,以及为什么它是研究活细胞中分子膜动力学的重要工具。与传统的 FCS 相比,STED-FCS 方法具有更好的可能性来区分自由扩散和异常扩散,从而深入了解脂质-蛋白相互作用和传统的脂质“筏”理论。

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