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3D correlative cryo-structured illumination fluorescence and soft X-ray microscopy elucidates reovirus intracellular release pathway

DOI: 10.1016/j.cell.2020.05.051 DOI Help

Authors: Ilias Kounatidis (Diamond Light Source) , Megan L. Stanifer (Heidelberg University Hospital) , Michael A. Phillips (Diamond Light Source; University of Oxford) , Perrine Paul-Gilloteaux (Université de Nantes, CNRS, INSERM) , Xavier Heiligenstein (CryoCapCell) , Hongchang Wang (Diamond Light Source) , Chidinma Okolo (Diamond Light Source) , Thomas M. Fish (Diamond Light Source) , Matthew C. Spink (Diamond Light Source) , David I. Stuart (University of Oxford) , Ilan Davis (University of Oxford) , Steeveh Boulant (Heidelberg University Hospital; German Cancer Research Center (DKFZ)) , Jonathan M. Grimes (Diamond Light Source; University of Oxford) , Ian M. Dobbie (University of Oxford) , Maria Harkiolaki (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Cell

State: Published (Approved)
Published: June 2020
Diamond Proposal Number(s): 21046 , 18314

Open Access Open Access

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.

Journal Keywords: correlative imaging; structured illumination microscopy; X-ray tomography; reovirus biology; CLXT

Diamond Keywords: Viruses

Subject Areas: Technique Development, Biology and Bio-materials


Instruments: B24-Cryo Soft X-ray Tomography

Discipline Tags:

Pathogens Infectious Diseases Health & Wellbeing Technique Development - Life Sciences & Biotech Life Sciences & Biotech

Technical Tags:

Microscopy X-ray Microscopy Correlative Light X-ray Microscopy (CLXM)