University of Leipzig 7 articles published in JoVE Biology MRM Microcoil Performance Calibration and Usage Demonstrated on Medicago truncatula Roots at 22 T Remco van Schadewijk1, Julia R. Krug2,3,4, Andrew Webb5, Henk Van As2,4, Aldrik H. Velders3,4, Huub J. M. de Groot1, A. Alia1,6 1Solid-state NMR, Leiden Institute of Chemistry, Faculty of Science, Leiden University, 2Laboratory of Biophysics, Wageningen University & Research, 3Laboratory of BioNanoTechnology, Wageningen University & Research, 4MAGNEtic resonance Facility, Wageningen University & Research, 5C.J. Gorter Center for High Field MRI, Radiology department, Leiden University Medical Centre, Leiden University, 6Institute for Medical Physics and Biophysics, Leipzig University A protocol to study biological tissue at high spatial resolution using ultra-high field magnetic resonance microscopy (MRM) using microcoils is presented. Step-by-step instructions are provided for characterizing the microcoils. Finally, optimization of imaging is demonstrated on plant roots. Chemistry Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells Robert Serfling*1, Lisa Seidel*1, Thore Böttke*1, Irene Coin1 1Institute of Biochemistry, Faculty of Life Sciences, University of Leipzig A facile fluorescence assay is presented to evaluate the efficiency of amino-acyl-tRNA-synthetase/tRNA pairs incorporating non-canonical amino-acids (ncAAs) into proteins expressed in mammalian cells. The application of ncAAs to study G-protein coupled receptors (GPCRs) is described, including photo-crosslinking mapping of binding sites and bioorthogonal GPCR labeling on live cells. Medicine Lavage-induced Surfactant Depletion in Pigs As a Model of the Acute Respiratory Distress Syndrome (ARDS) Martin Russ1, Sebastian Kronfeldt1, Willehad Boemke1, Thilo Busch2, Roland C. E. Francis*1, Philipp A. Pickerodt*1 1Department of Anesthesiology and Intensive Care Medicine, Campus Charité Mitte and Campus Virchow-Klinikum, Charité - Universitätsmedizin, Berlin, 2Department of Anesthesiology and Intensive Care Medicine, University of Leipzig Medical Faculty Repeated pulmonary lavages in anesthetized pigs induce lung injury resembling major aspects of human acute respiratory distress syndrome (ARDS). For this purpose the lungs are repeatedly lavaged with 0.9% saline at 37 °C. The goal of the protocol is a reproducible mitigation of gas exchange and hemodynamics for research in ARDS. Immunology and Infection Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples Jörg Flemmig1, Pauline Schwarz1, Ingo Bäcker2, Anna Leichsenring2, Franziska Lange2, Jürgen Arnhold1 1Institute for Medical Physics and Biophysics, Medical Faculty, University of Leipzig, 2Fraunhofer Institute for Cell Therapy and Immunology (IZI) Leipzig This protocol describes the quick enrichment of leukocytes from small blood samples for a subsequent specific determination of the halogenating peroxidase activity within the cells. The method can be applied to human and non-human material and may contribute to the evaluation of new inflammatory markers. Immunology and Infection Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain Claudia Pösel*1, Karoline Möller*1, Johannes Boltze1,2, Daniel-Christoph Wagner1, Gesa Weise1,3 1Ischemia Research Unit, Fraunhofer Institute for Cell Therapy and Immunology, 2Fraunhofer Research Institution for Marine Biotechnology, University of Lubeck, 3Department of Neurology, University of Leipzig Inflammation plays a central role in the pathogenesis of ischemic stroke. Increasing evidence suggests that it acts as a double-edged sword which exacerbates early brain injury, but also contributes to later repair. This protocol describes the isolation of immune cells from the ischemic brain and their subsequent flow cytometric phenotyping. Medicine Non-fluoroscopic Catheter Tracking for Fluoroscopy Reduction in Interventional Electrophysiology Philipp Sommer1, Simon Kircher1, Sascha Rolf1, Sergio Richter1, Micha Doering1, Arash Arya1, Andreas Bollmann1, Gerhard Hindricks1 1Department of Electrophysiology, Heart Center, University of Leipzig Radiation exposure is an underestimated risk in complex ablation procedures. Here, we describe a protocol to significantly decrease fluoroscopy time and dosage for both the patient and the lab staff by using a novel non-fluoroscopic catheter visualization system. Engineering High-Sensitivity Nuclear Magnetic Resonance at Giga-Pascal Pressures: A New Tool for Probing Electronic and Chemical Properties of Condensed Matter under Extreme Conditions Thomas Meier1, Jürgen Haase1 1Faculty of Physics and Earth Sciences, University of Leipzig Nuclear magnetic resonance is one of the most important spectroscopic tools. Here, the development of a new approach under high pressure, currently up to 10.1 GPa, is presented. This opens a new window into condensed matter physics and chemistry, where high-pressure research is of great importance.
