Charite---Universitatsmedizin-Berlin View Institution's Website 46 articles published in JoVE Medicine A Model of Reverse Vascular Remodeling in Pulmonary Hypertension Due to Left Heart Disease by Aortic Debanding in Rats Pengchao Sang1,2,3, Mariya M. Kucherenko1,2,3, Juquan Yao2, Qiuhua Li2, Szandor Simmons2,3, Wolfgang M. Kuebler*2,3, Christoph Knosalla*1,3,4 1Department of Cardiothoracic and Vascular Surgery, German Heart Center Berlin (DHZB), 2Institute of Physiology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 3DZHK (German Centre for Cardiovascular Research), Partner Site Berlin, 4Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health The present protocol describes a surgical procedure to remove ascending-aortic banding in a rat model of pulmonary hypertension due to left heart disease. This technique studies endogenous mechanisms of reverse remodeling in the pulmonary circulation and the right heart, thus informing strategies to reverse pulmonary hypertension and/or right ventricular dysfunction. Neuroscience Endovascular Perforation Model for Subarachnoid Hemorrhage Combined with Magnetic Resonance Imaging (MRI) Shuheng Liu*1, Katharina Tielking*1, Dario von Wedel1, Melina Nieminen-Kelhä1, Susanne Mueller2,3, Philipp Boehm-Sturm2,3, Peter Vajkoczy1, Ran Xu1 1Department of Neurosurgery, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, and Humboldt-Universität zu Berlin, and Berlin Institute of Health, 2Department of Experimental Neurology and Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 3NeuroCure Cluster of Excellence and Charité Core Facility 7T Experimental MRIs, Charité - Universitätsmedizin Berlin Here we present a standardized SAH mouse model, induced by endovascular filament perforation, combined with magnetic resonance imaging (MRI) 24 h after operation to ensure the correct bleeding site and exclude other relevant intracranial pathologies. Medicine An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery Julia A. Dubowitz1,2,3, Fabian Jost-Brinkmann1,4,5, Alexandra I. Ziegler1, Ryan D. Gillis1, Bernhard Riedel1,2,3,6, Erica K. Sloan1,2 1Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University, 2Department of Anaesthesia, Division of Cancer Surgery, Peter MacCallum Cancer Centre, 3Department of Critical Care, Melbourne Medical School, University of Melbourne, 4Medical Department, Division of Hepatology and Gastroenterology, Charité - Universitätsmedizin Berlin, 5Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 6Sir Peter MacCallum Department of Oncology, University of Melbourne This paper describes a method for modeling total intravenous anesthesia (TIVA) during cancer resection surgery in mice. The goal is to replicate key features of anesthesia delivery to patients with cancer. The method allows investigation of how anesthetic technique affects cancer recurrence after resection surgery. Immunology and Infection Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples Dario L. Frey*1,2, Matteo Guerra*1,2,3,4, Marcus A. Mall1,2,5,6,7, Carsten Schultz1,3,8 1Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), 2Dept. of Translational Pulmonology, University of Heidelberg, 3Molecular Medicine Partnership Unit (MMPU), European Molecular Biology Laboratory (EMBL), University of Heidelberg, 4Faculty of Biosciences, Collaboration for Joint Ph.D. Degree between EMBL and Heidelberg University, University of Heidelberg, 5Dept. of Pediatric Pulmonology, Immunology and Critical Care Medicine, Charité – Universitätsmedizin Berlin, 6Berlin Institute of Health (BIH), 7German Center for Lung Research (DZL), Associated Partner Site, Berlin, 8Dept. of Chemical Physiology and Biochemistry, Oregon Health and Science University The protocols herein described provide a guide to visualize and quantify the activity of neutrophil proteases in human sputum. The applications of such analysis span from the evaluation of anti-inflammatory treatments, to biomarker validation, drug screening and large cohort clinical studies. Medicine TBase - an Integrated Electronic Health Record and Research Database for Kidney Transplant Recipients Danilo Schmidt1, Bilgin Osmanodja2, Matthias Pfefferkorn2, Verena Graf2, Dirk Raschke1, Wiebke Duettmann2, Marcel G. Naik2, Carolin J. Gethmann2, Manuel Mayrdorfer2, Fabian Halleck2, Lutz Liefeldt2, Petra Glander2, Oliver Staeck2, Michael Mallach1, Martin Peuker1, Klemens Budde2 1Division IT, Charité - Universitätsmedizin Berlin, 2Department of Nephrology and Medical Intensive Care, Charité - Universitätsmedizin Berlin TBase combines an electronic health record with an innovative research database for kidney transplant recipients. TBase is built upon an in-memory database platform, connected to different hospital systems, and used for regular outpatient care. It automatically integrates all relevant clinical data including transplantation-specific data creating a unique research database. Medicine Digital Home-Monitoring of Patients after Kidney Transplantation: The MACCS Platform Wiebke Duettmann1, Marcel G. Naik1, Danilo Schmidt2, Matthias Pfefferkorn1, Matthias Kurz1, Verena Graf1, Andrea Kreichgauer3, Severin Hoegl4, Max Haenska4, Tim Gielsdorf1, Tillmann Breitenstein1, Bilgin Osmanodja1, Petra Glander1, Jannis Bakker1, Manuel Mayrdorfer1, Carolin J. Gethmann1, Friederike Bachmann1, Mira Choi1, Eva Schrezenmeier1, Bianca Zukunft1, Fabian Halleck1, Klemens Budde1 1Department of Nephrology and internal intensive Care, Charité Universitätsmedizin Berlin, 2IT Department, Charité Universitätsmedizin Berlin, 3Department of Neurology, Charité Universitätsmedizin Berlin, 4comjoodoc business solutions GmbH The MACCS platform is a comprehensive telemedicine concept aiming at better outcomes after kidney transplantation by sharing key medical information between patients and physicians. A telemedicine team reviews incoming data to detect potential complications and to improve adherence in kidney transplant recipients to achieve better long-term outcomes. Medicine Magnetic Resonance Imaging of Multiple Sclerosis at 7.0 Tesla Sonia Waiczies*1, Antje Els*1, Joseph Kuchling*2,3,4, Karin Markenroth Bloch5, Anna Pankowska6,7, Helmar Waiczies8, Carl Herrmann1, Claudia Chien2,3, Carsten Finke4,9, Friedemann Paul2,3,4, Thoralf Niendorf1,2,8 1Berlin Ultrahigh Field Facility (B.U.F.F.), Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 2Experimental and Clinical Research Center, Max Delbrueck Center for Molecular Medicine and Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 3NeuroCure Clinical Research Center, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 4Department of Neurology, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 5The Swedish National 7T Facility, Lund University Bioimaging Center, Lund University, 6Department of Radiography, Medical University of Lublin, 7ECOTECH-COMPLEX, Maria Curie-Skłodowska University, 8MRI.TOOLS GmbH, 9Berlin School of Mind and Brain, Humboldt-Universität zu Berlin Here, we present a protocol to acquire magnetic resonance (MR) images of multiple sclerosis (MS) patient brains at 7.0 Tesla. The protocol includes preparation of the setup including the radio-frequency coils, standardized interview procedures with MS patients, subject positioning in the MR scanner and MR data acquisition. Immunology and Infection Investigating Aortic Valve Calcification via Isolation and Culture of T Lymphocytes using Feeder Cells from Irradiated Buffy Coat Lavinia Curini*1,2, Mary Roxana Christopher*1, Herko Grubitzsch3,4, Ulf Landmesser1,3, Amedeo Amedei*2,5, Alexander Lauten*1,6, Brunilda Alushi*1,6 1Department of Cardiology, Campus Benjamin Franklin, Charité Universitätsmedizin Berlin and German Centre for Cardiovascular Research (DZHK), 2Department of Experimental and Clinical Medicine, University of Florence, 3Berlin Institute of Health, 4Department of Cardiology, German Heart Centre Berlin (DHZB), 5Sod of Interdisciplinary Internal Medicine, Azienda Ospedaliera Universitaria Careggi (AOUC), 6Department of General and Interventional Cardiology, Helios Klinikum Erfurt In this study, we describe the process of T lymphocyte isolation from fresh samples of calcified aortic valves and the analytical steps of T cell-cloning for the characterization of the adaptive leukocyte subsets by using flow cytometry analysis. Medicine Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry Wen-Jia Liu*1,2, Lisa Ernst*2, Benedict Doorschodt2, Jan Bednarsch1, Felix Becker3, Richi Nakatake2, Yuki Masano2, Ulf Peter Neumann1,4, Sven Arke Lang1, Peter Boor5, Isabella Lurje6, Georg Lurje1,6, René Tolba2, Zoltan Czigany1,2 1Department of Surgery and Transplantation, Faculty of Medicine, University Hospital RWTH Aachen, 2Institute for Laboratory Animal Science, Faculty of Medicine, University Hospital RWTH Aachen, 3Department of General-, Visceral-, and Transplantation Surgery, University Hospital Muenster, 4Department of Surgery, Maastricht University Medical Centre (MUMC), 5Institute of Pathology, Faculty of Medicine, University Hospital RWTH Aachen, 6Department of Surgery, Campus Charité Mitte | Campus Virchow-Klinikum, Charité-Universitätsmedizin Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions. Biology In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2 Maria Lucia Pigazzini1,2, Janine Kirstein1,3 1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund, 2NeuroCure Cluster of Excellence, Charité - Universitätsmedizin Berlin, 3Department of Biology and Chemistry, University of Bremen Presented here is a protocol to monitor degradation of the protein huntingtin fused to the photoconvertible fluorophore Dendra2. Neuroscience Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy Paul V. Ritschl*1,2,3, Lena Hofhansel*2,4, Bernhard Flörchinger5, Rupert Oberhuber2, Robert Öllinger1, Johann Pratschke1, Katja Kotsch6 1Department of Surgery Campus Charité Mitte/Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, 2Department of Visceral, Transplant and Thoracic Surgery, Medical University of Innsbruck, 3Charité Clinician Scientist Program, Berlin Institute of Health, 4Department of Psychiatry, Psychotherapy and Psychosomatics, RWTH Aachen University, 5Department of Cardiothoracic Surgery, University Medical Center Regensburg, 6Department of General, Visceral and Vascular Surgery, Charité - Universitätsmedizin Berlin We present a murine model of brain death induction in order to evaluate the influence of its pathophysiological effects on organs as well as on consecutive grafts in the context of solid organ transplantation. Biochemistry Characterization of Amyloid Structures in Aging C. Elegans Using Fluorescence Lifetime Imaging Maria Lucia Pigazzini*1,2, Christian Gallrein*1, Manuel Iburg*1, Gabriele Kaminski Schierle3, Janine Kirstein1,4 1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund Berlin, 2NeuroCure Cluster of Excellence, Charité - Universitätsmedizin Berlin, 3Molecular Neuroscience Group, Department of Chemical Engineering and Biotechnology, University of Cambridge, 4Cell Biology, University of Bremen Fluorescence lifetime imaging monitors, quantifies and distinguishes the aggregation tendencies of proteins in living, aging, and stressed C. elegans disease models. Neuroscience Single Synapse Indicators of Glutamate Release and Uptake in Acute Brain Slices from Normal and Huntington Mice Anton Dvorzhak1, Rosemarie Grantyn1 1Synaptic Dysfunction Group, Neuroscience Research Center, Charité - University Medicine We present a protocol to evaluate the balance between glutamate release and clearance at single corticostriatal glutamatergic synapses in acute slices from adult mice. This protocol uses the fluorescent sensor iGluu for glutamate detection, a sCMOS camera for signal acquisition and a device for focal laser illumination. Neuroscience Primary Cell Culture of Purified GABAergic or Glutamatergic Neurons Established through Fluorescence-activated Cell Sorting Paul Turko1, Keenan Groberman1, Toralf Kaiser2, Yuchio Yanagawa3, Imre Vida1 1Institut für Integrative Neuroanatomie, Charité, Universitätsmedizin Berlin, 2Department for Cytometry and Cellsorting, German Rheumatism Research Center, Leibniz Institute, 3Departments of Genetic and Behavioral Neuroscience, Graduate School of Medicine, Gunma University This protocol describes a cell sorting based method for the purification and culture of fluorescent GABAergic or glutamatergic neurons from the neocortex and hippocampus of postnatal mice or rats. Cancer Research Morphology-Based Distinction Between Healthy and Pathological Cells Utilizing Fourier Transforms and Self-Organizing Maps Fabian L. Kriegel1,2, Ralf Köhler2, Jannike Bayat-Sarmadi2, Simon Bayerl3, Anja E. Hauser2,3, Raluca Niesner2, Andreas Luch*1, Zoltan Cseresnyes*4 1Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), 2Deutsches Rheuma-Forschungszentrum (DRFZ) Berlin, a Leibniz Institute, 3Charité Universitätsmedizin Berlin, 4Applied Systems Biology, Leibniz Institute for Natural Product Research and Infection Biology Hans Knöll Institute Here, we provide a workflow that allows the identification of healthy and pathological cells based on their 3-dimensional shape. We describe the process of using 2D projection outlines based on the 3D surfaces to train a Self-Organizing Map that will provide objective clustering of the investigated cell populations. Medicine Predicting Treatment Response to Image-Guided Therapies Using Machine Learning: An Example for Trans-Arterial Treatment of Hepatocellular Carcinoma Aaron Abajian1, Nikitha Murali1, Lynn Jeanette Savic1,2, Fabian Max Laage-Gaupp1, Nariman Nezami1, James S. Duncan3, Todd Schlachter1, MingDe Lin4, Jean-François Geschwind5, Julius Chapiro1 1Department of Radiology and Biomedical Imaging, Yale School of Medicine, 2Department of Diagnostic and Interventional Radiology, Universitätsmedizin Charité Berlin, 3Department of Biomedical Engineering, Yale School of Engineering and Applied Science, 4Philips Research North America, 5Prescience Labs Intra-arterial therapies are the standard of care for patients with hepatocellular carcinoma who cannot undergo surgical resection. A method for predicting response to these therapies is proposed. The technique uses pre-procedural clinical, demographic, and imaging information to train machine learning models capable of predicting response prior to treatment. Medicine Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction David Bode1,2, Tim Guthof1, Burkert M. Pieske1,2, Frank R. Heinzel1,2, Felix Hohendanner1,2 1Department of Internal Medicine and Cardiology, Charité University Medicine, 2German Center for Cardiovascular Research (DZHK) Here, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes from a rat model of metabolic syndrome-related heart failure with preserved ejection fraction. A manual regulation of intraluminal pressure of cardiac cavities is implemented to yield functionally intact myocytes suitable for excitation-contraction-coupling studies. Bioengineering Metabolic Glycoengineering of Sialic Acid Using N-acyl-modified Mannosamines Paul R. Wratil1,2, Rüdiger Horstkorte3 1Max von Pettenkofer-Institut & Genzentrum, Virologie, Nationales Referenzzentrum für Retroviren, Medizinische Fakultät, LMU München, 2Institut für Laboratoriumsmedizin, klinische Chemie und Pathobiochemie, Charité - Universitätsmedizin Berlin, 3Institut für Physiologische Chemie, Martin-Luther-Universität Halle-Wittenberg Sialic acid is a typical monosaccharide-unit found in glycoconjugates. It is involved in a plethora of molecular and cellular interactions. Here we present a method to modify cell surface sialic acid expression using metabolic glycoengineering with N-acetylmannosamine derivatives. Biology A Protocol for Using Gene Set Enrichment Analysis to Identify the Appropriate Animal Model for Translational Research Christopher Weidner1, Matthias Steinfath1, Elisa Wistorf1, Michael Oelgeschläger1, Marlon R. Schneider1, Gilbert Schönfelder1,2 1Department of Experimental Toxicology and ZEBET, German Federal Institute for Risk Assessment (BfR), 2Department of Clinical Pharmacology and Toxicology, Charité-Universitätsmedizin Berlin We provide a standardized protocol for the use of gene set enrichment analysis of transcriptomic data to identify an ideal mouse model for translational research. This protocol can be used with DNA microarray and RNA sequencing data and can further be extended to other omics data if data are available. Neuroscience Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats Jens K. Haumesser1, Johanna Kühn1, Christopher Güttler1, Dieu-Huong Nguyen1, Maximilian H. Beck1, Andrea A. Kühn1, Christoph van Riesen1 1Department of Neurology, Movement Disorder and Neuromodulation Unit Berlin, Charité University Medicine Berlin In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia. Neuroscience Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice Simon H. Bayerl1, Melina Nieminen-Kelhä1, Thomas Broggini2, Peter Vajkoczy1, Vincent Prinz1 1Department of Neurosurgery and Center for Stroke-research Berlin (CSB), Charité-Universitätsmedizin, 2Department of Physics, University of California San Diego Surgical occlusion of a distal middle cerebral artery branch (MCAo) is a frequently used model in experimental stroke research. This manuscript describes the basic technique of permanent MCAo, combined with the insertion of a lateral cranial window, which offers the opportunity for longitudinal intravital microscopy in mice. 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. Behavior Measurement of Vibration Detection Threshold and Tactile Spatial Acuity in Human Subjects Rabih Moshourab1,2, Henning Frenzel2, Stefan Lechner3, Julia Haseleu2, Valérie Bégay2, Damir Omerbašić2, Gary R. Lewin2 1Department of Anesthesiology and Intensive Care Medicine, Charite Universitätsmedzin, Campus Virchow Klinikum und Campus Charite Mitte, 2Department of Neuroscience, Molecular Physiology of Somatic Sensation, Max Delbrück Center for Molecular Medicine, 3Institute of Pharmacology, University of Heidelberg Here, we present protocols to determine vibration detection thresholds and tactile acuity using psychophysical methods in man. Immunology and Infection A RAPID Method for Blood Processing to Increase the Yield of Plasma Peptide Levels in Human Blood Pauline Teuffel1, Miriam Goebel-Stengel1,2, Tobias Hofmann1, Philip Prinz1, Sophie Scharner1, Jan L. Körner3, Carsten Grötzinger3, Matthias Rose1, Burghard F. Klapp1, Andreas Stengel1 1Charité Center for Internal Medicine and Dermatology, Division General Internal and Psychosomatic Medicine, Charité Universitätsmedizin Berlin, Campus Benjamin Franklin, 2Department of Internal Medicine, Institute of Neurogastroenterology and Motility, Martin-Luther Hospital, Academic Teaching Institution of Charité Universitätsmedizin Berlin, 3Department of Hepatology and Gastroenterology, Molecular Cancer Research Center (MKFZ), Campus Virchow-Klinikum, Charité-Universitätsmedizin Berlin The RAPID blood processing method can be used in humans and yields higher peptide levels as well as allows for assessment of the correct molecular form. Therefore, this method will be a valuable tool in peptide research. Biology Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells Victoria Kegel*1, Daniela Deharde*1, Elisa Pfeiffer1, Katrin Zeilinger2, Daniel Seehofer1, Georg Damm1 1Department of General-, Visceral- and Transplantation Surgery, Charité University Medicine Berlin, 2Brandenburg Center for Regenerative Therapies, Charité University Medicine Berlin A technique to isolate human hepatocytes and non-parenchymal liver cells from the same donor is described. The different liver cell types build the basis for functional liver models and tissue engineering. This new method aims to isolate liver cells in a high yield and viability. Medicine Orthotopic Hind Limb Transplantation in the Mouse Georg J. Furtmüller*1, Byoungchol Oh*1, Johanna Grahammer*2, Cheng-Hung Lin3, Robert Sucher4, Madeline L. Fryer1, Giorgio Raimondi1, W.P. Andrew Lee1, Gerald Brandacher1 1Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine, 2Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, 3Center for Vascularized Composite Allotransplantation, Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital and School of Medicine, 4Department of General, Visceral and Transplant Surgery, Charite Berlin This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA). Bioengineering Procedure for Decellularization of Rat Livers in an Oscillating-pressure Perfusion Device Karl Hillebrandt1, Dietrich Polenz1, Antje Butter1, Peter Tang1, Anja Reutzel-Selke1, Andreas Andreou1, Hendrik Napierala1, Nathanael Raschzok1, Johann Pratschke1, Igor M. Sauer1, Benjamin Struecker1 1General, Visceral, and Transplantation Surgery, Charité – Universitätsmedizin Berlin The presented techniques for liver harvesting, cannulation and perfusion using our proprietary device enable sophisticated perfusion set-ups to improve decellularization and recellularization experiments in rat livers. Medicine Robotic Ablation of Atrial Fibrillation Alexander Wutzler1, Thomas Wolber2, Wilhelm Haverkamp1, Leif-Hendrik Boldt1 1Department of Cardiology, Charité — Universitätsmedizin Berlin, Campus Virchow, 2Department of Cardiology, University Hospital Zurich Pulmonary vein isolation (PVI) with an ablation catheter is a curative treatment for atrial fibrillation (AF). Robotic catheter systems aim to improve catheter steerability. Here, a procedure with a new robotic catheter system is presented. The goal of the procedure is electrical block between pulmonary vein and left atrium. Developmental Biology Automated Quantification of Hematopoietic Cell – Stromal Cell Interactions in Histological Images of Undecalcified Bone Sandra Zehentmeier1, Zoltan Cseresnyes2,3, Juan Escribano Navarro4, Raluca A. Niesner2, Anja E. Hauser1,5 1Immunodynamics, German Rheumatism Research Center, a Leibniz Institute, 2Biophysical Analytics, German Rheumatism Research Center, a Leibniz Institute, 3Max-Delbrück Center for Molecular Medicine, 4Wimasis GmbH, 5Immunodynamics and Intravital Imaging, Charité - University of Medicine A strategy to quantitatively analyze histological data in the bone marrow is presented. Confocal microscopy of fluorescently labeled cells in tissue sections results in 2-dimensional images, which are automatically analyzed. Co-localization analyses of different cell types are compared to data from simulated images, giving quantitative information about cellular interactions. Neuroscience Whole Mount Labeling of Cilia in the Main Olfactory System of Mice Sonja Oberland1,2, Eva Maria Neuhaus1,2 1Pharmacology and Toxicology, Universitaetsklinikum Jena, 2Charite-Universitaetsmedizin Berlin Cilia of olfactory sensory neurons contain proteins of the signal transduction cascade, but a detailed spatial analysis of their distribution is difficult in cryosections. We describe here an optimized approach for whole mount labeling and en face visualization of ciliary proteins. Neuroscience Whole-cell Patch-clamp Recordings from Morphologically- and Neurochemically-identified Hippocampal Interneurons Sam A. Booker1, Jie Song1, Imre Vida1 1Institute of Integrative Neuroanatomy, NeuroCure Cluster of Excellence, Charité Universitätmedizin Cortical networks are controlled by a small, but diverse set of inhibitory interneurons. Functional investigation of interneurons therefore requires targeted recording and rigorous identification. Described here is a combined approach involving whole-cell recordings from single or synaptically-coupled pairs of neurons with intracellular labeling, post-hoc morphological and immunocytochemical analysis. Behavior Transcranial Direct Current Stimulation and Simultaneous Functional Magnetic Resonance Imaging Marcus Meinzer1,2, Robert Lindenberg2, Robert Darkow2, Lena Ulm2, David Copland1, Agnes Flöel2 1Centre for Clinical Research, University of Queensland, 2Department of Neurology, NeuroCure Clinical Research Centre, and Centre for Stroke Research Berlin, Charité Universitätsmedizin Transcranial direct current stimulation (tDCS) is a noninvasive brain stimulation technique. It has successfully been used in basic research and clinical settings to modulate brain function in humans. This article describes the implementation of tDCS and simultaneous functional magnetic resonance imaging (fMRI), to investigate the neural basis of tDCS effects. Immunology and Infection Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers Zoltan Cseresnyes*1,2, Laura Oehme*3, Volker Andresen4, Anje Sporbert2, Anja E. Hauser*3,5, Raluca Niesner*1 1Biophysical Analytics, German Rheumatism Research Center, Leibniz Institute, 2Microscopy Core Facility, Max-Delbrück Center for Molecular Medicine, 3Immunodynamics, German Rheumatism Research Center, Leibniz Institute, 4LaVision Biotec GmbH, 5Immunodynamics and Intravital Imaging, Charité - University of Medicine High-resolution intravital imaging with enhanced contrast up to 120 µm depth in lymph nodes of adult mice is achieved by spatially modulating the excitation pattern of a multi-focal two-photon microscope. In 100 µm depth we measured resolutions of 487 nm (lateral) and 551 nm (axial), thus circumventing scattering and diffraction limits. Behavior Assessment of Age-related Changes in Cognitive Functions Using EmoCogMeter, a Novel Tablet-computer Based Approach Philipp Fuge*1,2, Simone Grimm*1,2,3,4, Anne Weigand1,2,3, Yan Fan1,3, Matti Gärtner1,3, Melanie Feeser1,3, Malek Bajbouj1,2,3 1Cluster of Excellence "Languages of Emotion", Freie Universität Berlin, 2Department of Psychiatry, Campus Benjamin Franklin, Charité Berlin, 3Dahlem Institute for Neuroimaging of Emotion, Freie Universität Berlin, 4Clinic for Affective Disorders and General Psychiatry, Psychiatric University Hospital Zurich We tested the usability of a tablet-computer-based application (EmoCogMeter) in investigating the effects of age on cognition. Results show an age-related cognitive decline, thereby proving the usability of our application. Findings underline the great clinical and practical potential of a tablet-based application for detection and monitoring of cognitive dysfunction. Medicine Prehospital Thrombolysis: A Manual from Berlin Martin Ebinger1,2,5, Sascha Lindenlaub1,3,5, Alexander Kunz1,2,5, Michal Rozanski2,5, Carolin Waldschmidt2,5, Joachim E. Weber2,5, Matthias Wendt2,5, Benjamin Winter1,2,5, Philipp A. Kellner4,5, Sabina Kaczmarek4,5, Matthias Endres1,2,5, Heinrich J. Audebert1,2,5 1Center for Stroke Research Berlin (CSB), Charité - Universitätsmedizin Berlin, 2Klinik und Hochschulambulanz für Neurologie, Charité - Universitätsmedizin Berlin, 3Medical School of the Universität Hamburg, Universitätsklinikum Hamburg - Eppendorf, 4Berliner Feuerwehr, 5STEMO-Consortium Identification of suspected stroke in the dispatch center of the Berlin Fire Brigade prompts the deployment of a CT-equipped ambulance. If ischemic stroke is confirmed and contraindications are excluded prehospital thrombolysis is applied. Medicine Technique and Considerations in the Use of 4x1 Ring High-definition Transcranial Direct Current Stimulation (HD-tDCS) Mauricio F. Villamar1,2, Magdalena Sarah Volz1,3, Marom Bikson4, Abhishek Datta1,4, Alexandre F. DaSilva*5, Felipe Fregni*1 1Laboratory of Neuromodulation, Department of Physical Medicine & Rehabilitation, Spaulding Rehabilitation Hospital and Massachusetts General Hospital, Harvard Medical School, 2School of Medicine, Pontifical Catholic University of Ecuador, 3Charité University Medicine Berlin, 4The City College of The City University of New York, 5Headache & Orofacial Pain Effort (H.O.P.E.), Biologic & Materials Sciences, School of Dentistry, University of Michigan High-definition transcranial direct current stimulation (HD-tDCS), with its 4x1-ring montage, is a noninvasive brain stimulation technique that combines both the neuromodulatory effects of conventional tDCS with increased focality. This article provides a systematic demonstration of the use of 4x1 HD-tDCS, and the considerations needed for safe and effective stimulation. Biology Culturing Primary Rat Inner Medullary Collecting Duct Cells Dörte Faust1, Andrea Geelhaar1, Beate Eisermann1, Jenny Eichhorst2, Burkhard Wiesner2, Walter Rosenthal1,3, Enno Klussmann1 1Anchored Signalling, Max-Delbrück-Center for Molecular Medicine, 2Leibniz Institute for Molecular Pharmacology (FMP), 3Charité University Medicine Berlin Arginine-vasopressin (AVP) controls fine-tuning of body water homeostasis through facilitating water reabsorption by renal principal cells. Here, we present a protocol for the cultivation of primary rat inner medullary collecting duct cells suitable for the elucidation of molecular mechanisms underlying AVP-mediated water reabsorption. Medicine Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia Lukas Julius Speetzen1, Matthias Endres1, Alexander Kunz1 1Department of Neurology, Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, Germany There is accumulating evidence, that ischemic preconditioning (PC) – a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice. Immunology and Infection Monitoring Dendritic Cell Migration using 19F / 1H Magnetic Resonance Imaging Helmar Waiczies1,2, Martin Guenther1,2, Julia Skodowski1,2, Stefano Lepore1,2, Andreas Pohlmann2, Thoralf Niendorf1,2, Sonia Waiczies1,2 1Experimental and Clinical Research Center, A joint cooperation between the Charité Medical Faculty and the Max Delbrück Center for Molecular Medicine, 2Berlin Ultrahigh Field Facility (B.U.F.F.), Max Delbrück Center for Molecular Medicine Tracking of cells using MRI has gained remarkable attention in the past years. This protocol describes the labeling of dendritic cells with fluorine (19F)-rich particles, the in vivo application of these cells, and monitoring the extent of their migration to the draining lymph node with 19F/1H MRI and 19F MRS. Medicine Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma Yvonne Welte1,2, Cathrin Davies1,3, Reinhold Schäfer1,4, Christian R.A. Regenbrecht1,3,4 1Institute of Pathology, Laboratory of Molecular Tumor Pathology, Charité - Universitätsmedizin Berlin, 2Institute for Chemistry and Biochemistry, Free University Berlin, 3Laboratory for Functional Genomics Charité (LFGC), Charité - Universitätsmedizin Berlin, 4Comprehensive Cancer Center Charité, Charité - Universitätsmedizin Berlin This article describes the preparation of freshly obtained melanoma tissue into primary cell cultures, and how to remove contaminations of erythrocytes and fibroblasts from the tumor cells. Finally, we describe how CD133+ putative melanoma stem cells are sorted from the CD133- bulk using Magnetic Activated Cell Sorting (MACS). Biology Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes Clemens Köhncke1,2, Ulrike Lisewski1, Leonhard Schleußner1, Carolin Gaertner1, Saskia Reichert1, Torsten K. Roepke1,3 1Experimental and Clinical Research Center (ECRC), Charité Medical Faculty and Max-Delbrück Center for Molecular Medicine (MDC), 2Medical Department, Division of Cardiology, Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, 3Medical Department, Division of Cardiology and Angiology, Campus Mitte, Charité - Universitätsmedizin Berlin Kv channel dysfunction is associated with cardiac arrhythmias. In order to study the molecular mechanisms that lead to such arrhythmias we utilize a systematic protocol for isolation of atrial and ventricular cardiomyocytes from Kv channel ancillary subunit knockout mice. Isolated cardiomyocytes can then immediately be used for cellular electrophysiological studies, biochemical or immunofluorescence (IF) assays. Medicine Driving Simulation in the Clinic: Testing Visual Exploratory Behavior in Daily Life Activities in Patients with Visual Field Defects Johanna Hamel1,2, Antje Kraft1, Sven Ohl3, Sophie De Beukelaer1, Heinrich J. Audebert1,2, Stephan A. Brandt1 1Department of Neurology, Universitätsmedizin Charité, 2Center for Stroke Research Berlin (CSB), Universitätsmedizin Charité, 3Berlin School of Mind and Brain, Humboldt Universität zu Berlin Patients with visual deficits after stroke report about different constraints in daily life most likely due to variable compensatory strategies, which are difficult to differentiate in clinical routine. We present a clinical set-up which allows measurement of different compensatory head- and eye-movement-strategies and evaluating their effects on driving performance. Neuroscience Electrode Positioning and Montage in Transcranial Direct Current Stimulation Alexandre F. DaSilva1, Magdalena Sarah Volz2,3, Marom Bikson4, Felipe Fregni2 1Headache & Orofacial Pain Effort (H.O.P.E.), Biologic & Material Sciences, School of Dentistry, University of Michigan, 2Laboratory of Neuromodulation, Department of Physical Medicine & Rehabilitation, Spaulding Rehabilitation Hospital and Massachusetts General Hospital, Harvard Medical School, 3Charité, University Medicine Berlin, 4Department of Biomedical Engineering, The City College of New York Transcranial direct current stimulation (tDCS) is an established technique to modulate cortical excitability1,2. It has been used as an investigative tool in neuroscience due to its effects on cortical plasticity, easy operation, and safe profile. One area that tDCS has been showing encouraging results is pain alleviation 3-5. Immunology and Infection IgY Technology: Extraction of Chicken Antibodies from Egg Yolk by Polyethylene Glycol (PEG) Precipitation Diana Pauly1, Pablo A. Chacana2, Esteban G. Calzado3, Björn Brembs4, Rüdiger Schade5 1Center for Biological Security, Robert Koch-Institute, 2CICVyA - INTA Castelar, Instituto de Virología, 3Center of Molecular Immunology, Ciudad de la Habana, Cuba, 4Department of Biology, Chemistry, Pharmacy, Institute of Biology-Neurobiology, Free University of Berlin, 5Institut of Pharmacology, Charité-University Medicine of Berlin This protocol describes in particular the extraction of total IgY from egg yolk by means of polyethylene glycol precipitation and gives general information about IgY technology. Medicine Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model Odilo Engel1, Sabine Kolodziej1, Ulrich Dirnagl1, Vincent Prinz1 1Department for Experimental Neurology, Center for Stroke Research Berlin, Charité Universitätsmedizin Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice. Biology Bacterial Delivery of RNAi Effectors: Transkingdom RNAi Hermann Lage1, Andrea Krühn1 1Institute of Pathology, Charité Campus Mitte For development of RNA interference (RNAi)-based therapies, a novel strategy was developed, transkingdom RNAi (tkRNAi). This technology uses non-pathogenic bacteria to produce and deliver therapeutic short hairpin RNA (shRNA) into target cells. Here, tkRNAi was successfully applied for reversal of classical ABCB1-mediated multidrug resistance (MDR) of cancer cells.