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Diamond Light Source, Harwell Science and Innovation Campus

3 ARTICLES PUBLISHED IN JoVE

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Biochemistry

Fixed Target Serial Data Collection at Diamond Light Source
Sam Horrell 1, Danny Axford 1, Nicholas E. Devenish 1, Ali Ebrahim 1, Michael A. Hough 2, Darren A. Sherrell 1,3, Selina L. S. Storm 1,4, Ivo Tews 5, Jonathan A. R. Worrall 2, Robin L. Owen 1
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.

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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. Gilchrist 3, M. Alejandra Carbajal 1, Roland A. Fleck 1
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.

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JoVE Journal

Single Particle Cryo-Electron Microscopy: From Sample to Structure
Joshua B.R. White 1, Daniel P. Maskell 1, Andrew Howe 2, Martin Harrow 2, Daniel K. Clare 2, C. Alistair Siebert 2, Emma L. Hesketh 1, Rebecca F. Thompson 1
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.

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