Accedi

Diamond Light Source, Harwell Science and Innovation Campus

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

Fixed Target Serial Data Collection at Diamond Light Source

Sam Horrell¹, Danny Axford¹, Nicholas E. Devenish¹, Ali Ebrahim¹, Michael A. Hough², Darren A. Sherrell^1,3, Selina L. S. Storm^1,4, Ivo Tews⁵, Jonathan A. R. Worrall², Robin L. Owen¹

¹Diamond Light Source, Harwell Science and Innovation Campus, ²School of Life Sciences, University of Essex, ³X-ray Science Division, Argonne National Laboratory, ⁴European Molecular Biology Laboratory, Hamburg Outstation c/o DESY, ⁵Biological Sciences, Institute for Life Sciences, University of Southampton

We present a comprehensive guide to fixed target sample preparation, data collection, and data processing for serial synchrotron crystallography at Diamond beamline I24.

Biochemistry

Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography

Claudine Bisson^1,2, Corey W. Hecksel^3,4, James B. Gilchrist³, M. Alejandra Carbajal¹, Roland A. Fleck¹

¹Centre for Ultrastructural Imaging, New Hunt’s House, Guy’s Campus, King’s College London, ²Department of Biological Science, Birkbeck College, University of London, ³Electron Bio-Imaging Centre, Diamond Light Source, Harwell Science and Innovation Campus, ⁴SLAC National Accelerator Laboratory, Stanford University

Using focused ion beam milling to produce vitreous on-grid lamellae from plunge frozen biological samples for cryo-electron tomography.

JoVE Journal

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Joshua B.R. White¹, Daniel P. Maskell¹, Andrew Howe², Martin Harrow², Daniel K. Clare², C. Alistair Siebert², Emma L. Hesketh¹, Rebecca F. Thompson¹

¹Astbury Centre Structural Molecular Biology, School Molecular and Cellular Biology, Faulty Biological Sciences, University of Leeds, ²Diamond Light Source, Harwell Science and Innovation Campus

Structure determination of macromolecular complexes using cryoEM has become routine for certain classes of proteins and complexes. Here, this pipeline is summarized (sample preparation, screening, data acquisition and processing) and readers are directed towards further detailed resources and variables that may be altered in the case of more challenging specimens.