University Medical Center Gottingen 4 articles published in JoVE Biology Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current Philipp Tomsits*1,2,3, Dominik Schüttler*1,2,3, Stefan Kääb1,2, Sebastian Clauss*1,2,3, Niels Voigt*4,5,6 1Department of Medicine I, University Hospital Munich, Campus Großhadern, Ludwig-Maximilians University Munich (LMU), 2Partner Site Munich, Munich Heart Alliance (MHA), DZHK (German Centre for Cardiovascular Research), 3Walter Brendel Center of Experimental Medicine, Ludwig-Maximilians University Munich (LMU), 4Institute of Pharmacology and Toxicology, University Medical Center Göttingen, 5Partner Site Göttingen, DZHK (German Centre for Cardiovascular Research), 6Cluster of Excellence "Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells" (MBExC), University of Göttingen Mouse models allow studying key mechanisms of arrhythmogenesis. For this purpose, high quality cardiomyocytes are necessary to perform patch-clamp measurements. Here, a method to isolate murine atrial and ventricular myocytes via retrograde enzyme-based Langendorff perfusion, which allows simultaneous measurements of calcium-transients and L-type calcium current, is described. Chemistry Setup of Capillary Electrophoresis-Inductively Coupled Plasma Mass Spectrometry (CE-ICP-MS) for Quantification of Iron Redox Species (Fe(II), Fe(III)) Bernhard Michalke1, Desiree Willkommen1,4, Vivek Venkataramani2,3 1Research Unit Analytical BioGeoChemistry, Helmholz Zentrum München-German Research Center for Environmental Health, 2Department of Hematology and Medical Oncology, University Medical Center Göttingen (UMG), 3Institute of Pathology, University Medical Center Göttingen (UMG), 4RECIPE Chemicals + Instruments GmbH This iron redox speciation method is based on capillary electrophoresis-inductively coupled plasma mass spectrometry with sample stacking combined with short analysis in one run. The method quickly analyzes and provides low limits of quantification for iron redox species across a diverse range of tissues and biofluid samples. Neuroscience Quantifying the Heterogeneous Distribution of a Synaptic Protein in the Mouse Brain Using Immunofluorescence Rebecca Wallrafen1, Thomas Dresbach1, Julio S. Viotti1 1Institute of Anatomy and Embryology, University Medical Center Göttingen Here, we describe a quantitative approach to determining the distribution of a synaptic protein relative to a marker protein using immunofluorescence staining, confocal microscopy, and computer-based analysis. Immunology and Infection Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags Kerstin Menck1, Daniel Behme1, Mathias Pantke1, Norbert Reiling2, Claudia Binder1, Tobias Pukrop1, Florian Klemm1 1Department of Hematology and Oncology, University Medical Center Göttingen, 2Microbial Interface Biology Group, Research Center Borstel We present a simple and efficient protocol for the generation of human macrophages. Buffy coats are processed by double density gradient centrifugation and isolated monocytes are then differentiated to macrophages in Teflon-coated cell culture bags. This maximizes macrophage yields and facilitates cell harvesting for subsequent experiments.
