Journal
METHODS
Volume 55, Issue 2, Pages 137-143Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2011.07.009
Keywords
Single-virus tracking; Real-time fluorescence microscopy; Virus endocytosis; DiD fluorescent labeling; Virus fusion; Virus binding
Funding
- Pediatric Dengue Vaccine Initiative, The Netherlands Organization for Scientific Research Earth and Life Sciences (NWO-ALW)
- Erasmus Mundus External Cooperation Window
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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.
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