Ewers Helge, Schelhaas Mario
Institute of Biochemistry, ETH Zurich, Zurich, Switzerland.
Methods Enzymol. 2012;506:63-80. doi: 10.1016/B978-0-12-391856-7.00028-7.
Viruses have evolved to mimic cellular ligands in order to gain access to their host cells for replication. Since viruses are simple in structure, they rely on host cells for all their transportation needs. Following single virus particles during the initial phase of infection, that is, virus entry into target cells, can reveal crucial information on the mechanism of pathogen infections and likewise cellular transport and membrane dynamics. Here, we give an overview on how to fluorescently label virus particles for live cell microscopy, and on how virus entry can be analyzed by single particle tracking experiments. Highlighted are strategies, on how to chemically introduce fluorophores into virions, and on how to extract quantitative information from live cell data.
病毒已经进化到能够模拟细胞配体,以便进入宿主细胞进行复制。由于病毒结构简单,它们的所有运输需求都依赖于宿主细胞。在感染的初始阶段追踪单个病毒颗粒,即病毒进入靶细胞的过程,可以揭示有关病原体感染机制以及细胞运输和膜动力学的关键信息。在这里,我们概述了如何对病毒颗粒进行荧光标记以用于活细胞显微镜观察,以及如何通过单颗粒追踪实验分析病毒进入过程。重点介绍了将荧光团化学引入病毒粒子的策略,以及如何从活细胞数据中提取定量信息。