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Correlative Light and Electron Microscopy of Influenza Virus Entry and Budding

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Influenza Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1836))

Abstract

Influenza A virus (IAV) entry is a stepwise process regulated by viral and cellular cues, facilitating cellular functions. Virus entry begins by attachment of hemagglutinin to cell surface sialic acids, followed by endocytic uptake, vesicular transport along microtubules, low-pH-mediated viral membrane fusion with the late endosomal membrane, capsid uncoating, viral ribonucleoprotein (vRNP) release, and nuclear import of vRNPs. Here we show a basic methodology to visualize incoming and egressing IAV particles by correlative light and electron microscopy (CLEM). We combine fluorescence microscopy of virus-infected human lung carcinoma A549 cells with high-pressure freezing (HPF) and in-resin fluorescence CLEM and the Tokuyasu CLEM method. This approach forms a basis to study the virus life cycle and virus-host interactions at the ultrastructural level.

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Author information

Authors and Affiliations

  1. School of Biochemistry, University of Bristol, Bristol, UK

    Lorna Hodgson & Paul Verkade

  2. Wolfson Bioimaging Facility, University of Bristol, Bristol, UK

    Paul Verkade

  3. School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK

    Yohei Yamauchi

Authors
  1. Lorna Hodgson
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  2. Paul Verkade
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  3. Yohei Yamauchi
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Corresponding author

Correspondence to Yohei Yamauchi .

Editor information

Editors and Affiliations

  1. School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK

    Yohei Yamauchi

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Hodgson, L., Verkade, P., Yamauchi, Y. (2018). Correlative Light and Electron Microscopy of Influenza Virus Entry and Budding. In: Yamauchi, Y. (eds) Influenza Virus. Methods in Molecular Biology, vol 1836. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8678-1_12

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  • DOI: https://doi.org/10.1007/978-1-4939-8678-1_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8677-4

  • Online ISBN: 978-1-4939-8678-1

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