Correlative Light and Electron Microscopy of Influenza Virus Entry and Budding: Influenza Virus

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

147 Downloads (Pure)

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.
Original languageEnglish
Title of host publicationInfluenza Virus
Subtitle of host publicationMethods and Protocols
EditorsYohei Yamauchi
PublisherHumana Press
Chapter12
Pages237-260
Number of pages24
ISBN (Electronic)9781493986781
ISBN (Print)9781493986774
DOIs
Publication statusPublished - 28 Aug 2018

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume1836
ISSN (Print)1064-3745

Keywords

  • Influenza virus entry
  • Fluorescent confocal microscopy
  • Electron microscopy
  • CLEM
  • Tokuyasu
  • High-pressure freezing
  • Endosome

Fingerprint Dive into the research topics of 'Correlative Light and Electron Microscopy of Influenza Virus Entry and Budding: Influenza Virus'. Together they form a unique fingerprint.

  • Equipment

  • Cite this

    Hodgson, L., Verkade, P., & Yamauchi, Y. (2018). Correlative Light and Electron Microscopy of Influenza Virus Entry and Budding: Influenza Virus. In Y. Yamauchi (Ed.), Influenza Virus: Methods and Protocols (pp. 237-260). (Methods in Molecular Biology; Vol. 1836). Humana Press. https://doi.org/10.1007/978-1-4939-8678-1_12