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Deciphering Virus Entry with Fluorescently Labeled Viral Particles

  • Anja B. Hoffmann
  • Magalie Mazelier
  • Psylvia Léger
  • Pierre-Yves Lozach
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1836)

Abstract

To infect host cells, viruses have to gain access to the intracellular compartment. The infection process starts with the attachment of viruses to the cell surface. Then a complex series of events, highly dynamic, tightly intricate, and often hard to investigate, follows. This includes virus displacement at the plasma membrane, binding to receptors, signaling, internalization, and release of the viral genome and material into the cytosol. In the past decades, the emergence of sensitive, accurate fluorescence-based technologies has opened new perspectives of investigations in the field. Visualization of single viral particles in fixed and living cells as well as quantification of each virus entry step has been made possible. Here we describe the procedure to fluorescently label viral particles. We also illustrate how to use this powerful tool to decipher the entry of viruses with the most recent fluorescence-based techniques such as high-speed confocal and total internal reflection microscopy, flow cytometry, and fluorimetry.

Key words

Endocytosis Flow cytometry Fluorescent dyes Fluorescently labeled viral particles Fluorimetry Intracellular trafficking Membrane fusion Microscopy Single viral particle tracking Virus entry 

Notes

Acknowledgments

This work was supported by grants from CellNetworks Research Group funds and from DFG LO-2338/1-1.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Anja B. Hoffmann
    • 1
  • Magalie Mazelier
    • 1
  • Psylvia Léger
    • 1
  • Pierre-Yves Lozach
    • 1
  1. 1.From CellNetworks Cluster of Excellence and Department of Infectious Diseases, VirologyUniversity Hospital HeidelbergHeidelbergGermany

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