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3 ARTICLES PUBLISHED IN JoVE

Biology

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography

Johannes Groen¹, Andrea Sorrentino¹, Lucía Aballe¹, Robert Oliete¹, Ricardo Valcárcel¹, Chidinma Okolo², Ilias Kounatidis², Maria Harkiolaki², Ana Joaquina Pérez-Berná¹, Eva Pereiro¹

¹MISTRAL beamline, Alba Light Source, Cerdanyola del Valles, Spain, ²Beamline B24, Diamond Light Source, Harwell Science and Innovation Campus, Didcot, UK

Here, a protocol describing the sample preparation and data collection steps required in cryo soft X-ray tomography (SXT) to image the ultrastructure of whole cryo-preserved cells at a resolution of 25 nm half pitch, is presented.

Biology

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

Nina Vyas^*1, Nina Perry^*1, Chidinma A. Okolo¹, Ilias Kounatidis¹, Thomas M. Fish¹, Kamal L. Nahas^1,2, Archana Jadhav¹, Mohamed A. Koronfel¹, Johannes Groen³, Eva Pereiro³, Ian M. Dobbie⁴, Maria Harkiolaki¹

¹Harwell Science and Innovation Campus, Beamline B24, Diamond Light Source, ²Division of Virology, Department of Pathology, University of Cambridge, ³ALBA Synchrotron, Beamline 09 - MISTRAL, ⁴Micron Advanced Imaging Consortium, Department of Biochemistry, University of Oxford

This protocol demonstrates how to image biological cryo-preserved samples using cryo-structured illumination microscopy. We demonstrate the methodology by imaging the cytoskeleton of U2OS cells.

iNEXT-Discovery and Instruct-ERIC: Integrating High-End Services for Translational Research in Structural Biology

Hans Wienk¹, Lucia Banci², Susan Daenke³, Eva Pereiro⁴, Harald Schwalbe⁵, David Ian Stuart⁶, Manfred S. Weiss⁷, Anastassis Perrakis¹

¹Division of Biochemistry and Oncode Institute, The Netherlands Cancer Institute, Amsterdam, ²CERM/CIRMMP, University of Florence, ³Instruct-ERIC, Oxford University, ⁴MISTRAL beamline, Alba Light Source, ⁵Center for Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, ⁶Division of Structural Biology, Oxford University, ⁷Macromolecular Crystallographyy, Helmholtz-Zentrum Berlin

iNEXT-Discovery and Instruct-ERIC: Integrating High-End Services for Translational Research in Structural Biology