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Live Imaging of Influenza Viral Ribonucleoproteins Using Light-Sheet Microscopy

  • Amar R. Bhagwat
  • Valerie Le Sage
  • Seema S. LakdawalaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1836)

Abstract

Influenza viruses exhibit a complex life cycle that is still poorly understood. It involves independent replication of each of the eight segments that make up its genome and subsequent coordinated assembly as they egress from the host cell. Fast, time-resolved volumetric live cell imaging offers a powerful tool for understanding the various host mechanisms hijacked by the virus. Here, we describe the methods necessary for generating influenza viruses that carry a fluorescently tagged polymerase complex, infection of biologically relevant cells with these viruses, and finally protocols for live cell imaging and analysis.

Key words

Influenza genome Fluorescent influenza virus Fluorescence imaging Live cell imaging Particle tracking Fluorescence microscopy Light-sheet microscopy Fluorescent polymerase 

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Copyright information

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

Authors and Affiliations

  • Amar R. Bhagwat
    • 1
  • Valerie Le Sage
    • 1
  • Seema S. Lakdawala
    • 1
    Email author
  1. 1.Department of Microbiology and Molecular GeneticsUniversity of Pittsburgh School of MedicinePittsburghUSA

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