European Synchrotron Radiation Facility View Institution's Website 4 articles published in JoVE Biochemistry Fully Autonomous Characterization and Data Collection from Crystals of Biological Macromolecules Stephanie Hutin1, Bart Van Laer1, Christoph Mueller-Dieckmann1, Gordon Leonard1, Didier Nurizzo1, Matthew W. Bowler2 1Structural Biology Group, European Synchrotron Radiation Facility, 2Grenoble Outstation, European Molecular Biology Laboratory Here, we describe how to use the automated screening and data collection options available at some synchrotron beamlines. Scientists send cryocooled samples to the synchrotron, and the diffraction properties are screened, the data sets are collected and processed and, where possible, a structure solution is carried out—all without human intervention. Biochemistry Structure Solution of the Fluorescent Protein Cerulean Using MeshAndCollect Stephanie Hutin*1, Gianluca Santoni*1, Ulrich Zander2, Nicolas Foos1, Sylvain Aumonier1, Guillaume Gotthard1, Antoine Royant1,3, Christoph Mueller-Dieckmann1, Gordon Leonard1 1European Synchrotron Radiation Facility, Structural Biology Group, 2European Molecular Biology Laboratory, 3Univ. Grenoble Alpes, CNRS, CEA, IBS (Institut de Biologie Structurale) We present the use of the MeshAndCollect protocol to obtain a complete diffraction data set, for use in subsequent structure determination, composed of partial diffraction data sets collected from many small crystals of the fluorescent protein Cerulean. Biochemistry Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline Martha E. Brennich1, Adam R. Round2,3, Stephanie Hutin4 1Structural Biology Group, European Synchrotron Radiation Facility, 2European Molecular Biology Laboratory, 3School of Chemical and Physical Sciences, Keele University, 4Groupe de Microscopie Electronique et Méthodes, Institut de Biologie Structurale The determination of the solution structure of a protein by small angle X-ray scattering (SAXS) requires monodisperse samples. Here, we present two possibilities to ensure minimal delays between sample preparation and data acquisition: online size-exclusion chromatography (SEC) and online ion-exchange chromatography (IEC). Biology Non-invasive 3D-Visualization with Sub-micron Resolution Using Synchrotron-X-ray-tomography Michael Heethoff1, Lukas Helfen2, Peter Cloetens2 1Department of Evolutionary Biology of Invertebrates, University of Tubingen, 2European Synchrotron Radiation Facility We used synchrotron X-ray tomography at the European Synchrotron Radiation Facility (ESRF) to non-invasively produce 3D tomographic datasets with a pixel-resolution of 0.7µm. Using volume rendering software, this allows the reconstruction of internal structures in their natural state without the artefacts produced by histological sectioning.
