3D Correlative Cryo-Structured Illumination Fluorescence and Soft X-ray Microscopy Elucidates Reovirus Intracellular Release Pathway

Kounatidis, Ilias; Megan, Stanifer; Michael, Philips; Perrine, Paul-Gilloteaux; Xavier, Heiligenstein; Hongchang, Wang; Chidinma, Okolo; Thomas, Fish; Matthew, Spink; David, Stuart; Ilan, Davis; Steeve, Boulant; Jonathan, Grimes; Ian, Dobbie and Maria, Harkiolaki (2020). 3D Correlative Cryo-Structured Illumination Fluorescence and Soft X-ray Microscopy Elucidates Reovirus Intracellular Release Pathway. Cell, 182(2) pp. 515–530.

DOI: https://doi.org/10.1016/j.cell.2020.05.051

Abstract

Imaging of biological matter across resolution scales entails the challenge of preserving the direct and unambiguous correlation of subject features from the macroscopic to the microscopic level. Here, we present a correlative imaging platform developed specifically for imaging cells in 3D under cryogenic conditions by using X-rays and visible light. Rapid cryo-preservation of biological specimens is the current gold standard in sample preparation for ultrastructural analysis in X-ray imaging. However, cryogenic fluorescence localization methods are, in their majority, diffraction-limited and fail to deliver matching resolution. We addressed this technological gap by developing an integrated, user-friendly platform for 3D correlative imaging of cells in vitreous ice by using super-resolution structured illumination microscopy in conjunction with soft X-ray tomography. The power of this approach is demonstrated by studying the process of reovirus release from intracellular vesicles during the early stages of infection and identifying intracellular virus-induced structures.

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About

  • Item ORO ID
  • 85726
  • Item Type
  • Journal Item
  • ISSN
  • 1097-4172
  • Project Funding Details
  • Funded Project NameProject IDFunding Body
    Wellcome Centre for Human Genetics105605/Z/14/ZWelcome Trust
    Wellcome Trust ISSF203141/Z/16/ZWelcome Trust
    Welcome Trust Stategic Award091911Welcome Trust
    Welcome Trust Stategic Award107457Welcome Trust
    CROCOVALANR-18-CE45-0015France BioImaging
    Wellcome Investigator Award200835/Z/16/ZWelcome Trust
    Not SetBO 4340/1-1Heisenberg
    Not SetSFB1129Heisenberg
    Not Set240245660Deutsche Forschungsgemeinschaft
    Not SetMR/N00065X/1MRC
    B24 ProposalMX21046Diamond Light Source
    b24 ProposalMX18314Diamond Light Source
  • Keywords
  • Correlative Imaging; Structured Illumination Microscopy; X-ray Tomography; Reovirus Biology; CLXT; Virology
  • Academic Unit or School
  • Faculty of Science, Technology, Engineering and Mathematics (STEM) > Life, Health and Chemical Sciences
    Faculty of Science, Technology, Engineering and Mathematics (STEM)
  • Copyright Holders
  • © 2020 The Authors
  • Related URLs
  • Depositing User
  • Ilias Kounatidis

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