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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| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
23 July 2020
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| In: |
Cell
Year: 2020, Volume: 182, Issue: 2, Pages: 515-530.e17 |
| ISSN: | 1097-4172 |
| DOI: | 10.1016/j.cell.2020.05.051 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.cell.2020.05.051 Verlag, lizenzpflichtig, Volltext: http://www.sciencedirect.com/science/article/pii/S009286742030684X 
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| Author Notes: | Ilias Kounatidis, Megan L. Stanifer, Michael A. Phillips, Perrine Paul-Gilloteaux, Xavier Heiligenstein, Hongchang Wang, Chidinma A. Okolo, Thomas M. Fish, Matthew C. Spink, David I. Stuart, Ilan Davis, Steeve Boulant, Jonathan M. Grimes, Ian M. Dobbie, and Maria Harkiolaki |
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| 245 | 1 | 0 | |a 3D correlative cryo-structured illumination fluorescence and soft X-ray microscopy elucidates reovirus intracellular release pathway |c Ilias Kounatidis, Megan L. Stanifer, Michael A. Phillips, Perrine Paul-Gilloteaux, Xavier Heiligenstein, Hongchang Wang, Chidinma A. Okolo, Thomas M. Fish, Matthew C. Spink, David I. Stuart, Ilan Davis, Steeve Boulant, Jonathan M. Grimes, Ian M. Dobbie, and Maria Harkiolaki |
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| 520 | |a 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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