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

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 cry...

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Bibliographic Details
Published in:Cell Vol. 182; no. 2; p. 515
Main Authors: Kounatidis, Ilias, Stanifer, Megan L, Phillips, Michael A, Paul-Gilloteaux, Perrine, Heiligenstein, Xavier, Wang, Hongchang, Okolo, Chidinma A, Fish, Thomas M, Spink, Matthew C, Stuart, David I, Davis, Ilan, Boulant, Steeve, Grimes, Jonathan M, Dobbie, Ian M, Harkiolaki, Maria
Format: Journal Article
Language:English
Published: United States 23.07.2020
Subjects:
Cell Line, Tumor
Cryoelectron Microscopy - instrumentation
Cryoelectron Microscopy - methods
Endosomes - metabolism
Endosomes - virology
Fluorescent Dyes - chemistry
Humans
Imaging, Three-Dimensional
Microscopy, Fluorescence
Reoviridae - chemistry
Reoviridae - physiology
Virus Release - physiology
X-ray tomography
reovirus biology
CLXT
structured illumination microscopy
correlative imaging
ISSN:1097-4172, 1097-4172
Online Access:Get more information
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Summary: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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ISSN:1097-4172
1097-4172
DOI:10.1016/j.cell.2020.05.051