Alba Light Source, Cerdanyola del Valles, Spain

2 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.