Diamond Light Source, Harwell Science and Innovation Campus 4 articles published in JoVE Biochemistry Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography Claudine Bisson1,2, Corey W. Hecksel3,4, James B. Gilchrist3, M. Alejandra Carbajal1, Roland A. Fleck1 1 Using focused ion beam milling to produce vitreous on-grid lamellae from plunge frozen biological samples for cryo-electron tomography. Biochemistry 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. Chemistry Achieving Efficient Fragment Screening at XChem Facility at Diamond Light Source Alice Douangamath*1,2, Ailsa Powell*1,2, Daren Fearon*1,2, Patrick M. Collins1,2, Romain Talon1,2,3, Tobias Krojer3,4, Rachael Skyner1,2, Jose Brandao-Neto1,2, Louise Dunnett1,2, Alexandre Dias1,2, Anthony Aimon1,2,3, Nicholas M. Pearce1,3, Conor Wild3,5, Tyler Gorrie-Stone1, Frank von Delft1,2,3,4,6 1Diamond Light Source Ltd, Harwell Science and Innovation Campus, 2Research Complex at Harwell, Harwell Science and Innovation Campus, 3Structural Genomics Consortium, University of Oxford, 4Centre for Medicines Discovery, University of Oxford, 5Oxford Protein Informatics Group, Department of Statistics, Oxford University, 6Department of Biochemistry, University of Johannesburg This paper describes the complete XChem process for crystal-based fragment screening, starting from applying for access and all subsequent steps to data dissemination. Biochemistry Sample Preparation and Transfer Protocol for In-Vacuum Long-Wavelength Crystallography on Beamline I23 at Diamond Light Source Ramona Duman1, Christian M. Orr1, Vitaliy Mykhaylyk1, Kamel El Omari1, Robert Pocock1, Vinay Grama1, Armin Wagner1 1Diamond Light Source Ltd, Harwell Science & Innovation Campus Here, we present a protocol for cryogenic sample preparation and transfer of crystals into the vacuum endstation on beamline I23 at Diamond Light Source, for long-wavelength macromolecular X-ray crystallography experiments.
