Monitoring virus entry into living cells using DiD-labeled dengue virus particles

被引：33

作者：

Ayala-Nunez, Nilda V. ^{[1
]}

Wilschut, Jan ^{[1
]}

Smit, Jolanda M. ^{[1
]}

机构：

[1] Univ Groningen, Univ Med Ctr Groningen, Mol Virol Sect, Dept Med Microbiol, NL-9700 RB Groningen, Netherlands

来源：

METHODS | 2011年 / 55卷 / 02期

关键词：

Single-virus tracking; Real-time fluorescence microscopy; Virus endocytosis; DiD fluorescent labeling; Virus fusion; Virus binding; MEMBRANE-FUSION; IN-VITRO; ENDOCYTOSIS; TRACKING; ACIDIFICATION; MICROSCOPY; MECHANISM; INHIBITORS; INFECTION; ANALOGS;

D O I：

10.1016/j.ymeth.2011.07.009

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A variety of approaches can be applied to investigate the multiple steps and interactions that occur during virus entry into the host cell. Single-virus tracking is a powerful real-time imaging technique that offers the possibility to monitor virus-cell binding, internalization, intracellular trafficking behavior, and the moment of membrane fusion of single virus particles in living cells. Here we describe the development and applications of a single-virus tracking assay based on the use of DiD-labeled dengue virus (DENV) in BS-C-1 cells. In addition - and using the same experimental setup - we present a binding and fusion assay that can be used to obtain a rapid insight into the relative extent of virus binding to the cell surface and membrane fusion. Details of virus labeling and characterization, microscopy setup, protocols, data analysis, and hints for troubleshooting are described throughout the paper. (C) 2011 Elsevier Inc. All rights reserved.

引用

页码：137 / 143

页数：7

共 34 条

[1] Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrinmediated endocytosis [J].