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Instruments / Technical Area	Publication Details	Published	Actions
B24-Cryo Soft X-ray Tomography	3D correlative cryo-structured illumination fluorescence and soft X-ray microscopy elucidates reovirus intracellular release pathway Ilias Kounatidis , Megan L. Stanifer , Michael A. Phillips , Perrine Paul-Gilloteaux , Xavier Heiligenstein , Hongchang Wang , Chidinma Okolo , Thomas M. Fish , Matthew C. Spink , David I. Stuart , Ilan Davis , Steeveh Boulant , Jonathan M. Grimes , Ian M. Dobbie , Maria Harkiolaki Cell DOI: 10.1016/j.cell.2020.05.051 Diamond Proposal Number(s): [21046, 18314] Open Access Show Abstract ▼ 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.	Jun 2020

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B24-Cryo Soft X-ray Tomography

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