کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1993767 | 1064705 | 2011 | 7 صفحه PDF | دانلود رایگان |

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.
► We describe a single-virus tracking assay using DiD-labeled dengue virus.
► Single-virus tracking is used to monitor – in real-time – virus entry to the cell.
► We describe microscopy setup, protocols, data analysis, and troubleshooting.
► A binding and fusion assay using DiD-labeled virus is included.
► This assay is used to analyze virus binding to cell surface and membrane fusion.
Journal: Methods - Volume 55, Issue 2, October 2011, Pages 137–143