Otto-von-Guericke University Magdeburg View Institution's Website 15 articles published in JoVE Biochemistry Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex Laura K. Hillert-Richter1, Inna N. Lavrik1 1Translational Inflammation Research, Center of Dynamic Systems, Otto von Guericke University Magdeburg Here, an experimental workflow is presented that enables the detection of caspase-8 processing directly at the death-inducing signaling complex (DISC) and determines the composition of this complex. This methodology has broad applications, from unraveling the molecular mechanisms of cell death pathways to the dynamic modeling of apoptosis networks. Cancer Research Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo Julia Volckmar1,2, Laura Knop1,2,3, Tatjana Hirsch1, Sarah Frentzel1,2, Christian Erck4, Marco van Ham4, Sabine Stegemann-Koniszewski*1,2,5, Dunja Bruder*1,2 1Immune Regulation Group, Helmholtz Centre for Infection Research, 2Infection Immunology Group, Institute of Medical Microbiology, Infection Prevention and Control, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg, 3Institute for Molecular and Clinical Immunology, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg, 4Cellular Proteome Research, Helmholtz Centre for Infection Research, 5Experimental Pneumology, University Hospital of Pneumology, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg We describe a protocol for the chemical conjugation of the model antigen ovalbumin to an endocytosis receptor-specific antibody for in vivo dendritic cell targeting. The protocol includes purification of the antibody, chemical conjugation of the antigen, as well as purification of the conjugate and the verification of efficient conjugation. Engineering Evaluating Usability Aspects of a Mixed Reality Solution for Immersive Analytics in Industry 4.0 Scenarios Burkhard Hoppenstedt1, Thomas Probst2, Manfred Reichert1, Winfried Schlee3, Klaus Kammerer1, Myra Spiliopoulou4, Johannes Schobel1, Michael Winter1, Anna Felnhofer5, Oswald D. Kothgassner6, Rüdiger Pryss7 1Institute of Databases and Information Systems, Ulm University, 2Department for Psychotherapy and Biopsychosocial Health, Danube University Krems, 3Department of Psychiatry and Psychotherapy, University of Regensburg, 4Faculty of Computer Science, Otto von Guericke University Magdeburg, 5Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, 6Department of Child and Adolescent Psychiatry, Medical University of Vienna, 7Institute of Clinical Epidemiology and Biometry, University of Würzburg This protocol delineates the technical setting of a developed mixed reality application that is used for immersive analytics. Based on this, measures are presented, which were used in a study to gain insights into usability aspects of the developed technical solution. Medicine An In Vitro Hemodynamic Loop Model to Investigate the Hemocytocompatibility and Host Cell Activation of Vascular Medical Devices Max Wacker1, Ulf Betke2, Katrin Borucki3, Jörn Hülsmann1, George Awad1, Sam Varghese1, Maximilian Scherner1, Michael Hansen4, Jens Wippermann1, Priya Veluswamy1 1Department of Cardiothoracic Surgery, Otto-von-Guericke-University, 2Department of Mechanical Engineering, Institute for Materials and Joining Technology, Otto-von-Guericke-University, 3Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University, 4Division of Cardiology and Angiology, Department of Internal Medicine, Otto-von-Guericke-University Presented here is a protocol for a standardized in vitro hemodynamic loop model. This model allows to test the hemocompatibility of perfusion tubes or vascular stents to be in accordance with ISO (International Organization for Standardization) standard 10993-4. Environment A Guide to Concentration Alternating Frequency Response Analysis of Fuel Cells Antonio Sorrentino1, Kai Sundmacher1,2, Tanja Vidaković-Koch1 1Max Planck Institute for Dynamics of Complex Technical Systems, 2Process Systems Engineering, Otto-von-Guericke University Magdeburg We present a protocol for concentration alternating frequency response analysis of fuel cells, a promising new method of studying fuel cell dynamics. Neuroscience An Unbiased Approach of Sampling TEM Sections in Neuroscience Stefan Wernitznig1, Florian Reichmann2, Mariella Sele1, Christoph Birkl3, Johannes Haybäck4,5, Florian Kleinegger4, Anna Birkl-Töglhofer4, Stefanie Krassnig4, Christina Wodlej4, Peter Holzer2, Daniel Kummer1, Elisabeth Bock1, Gerd Leitinger1 1Department of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of Graz, 2Department of Pharmacology, Otto Loewi Research Center, Medical University of Graz, 3Division of General Neurology, Department of Neurology, Medical University of Graz, 4Institute of Pathology, Medical University of Graz, 5Department of Pathology, Medical Faculty, Otto von Guericke University Magdeburg We introduce a novel workflow for electron microscopy investigations of brain tissue. The method allows the user to examine neuronal features in an unbiased fashion. For elemental analysis, we also present a script that automatizes most of the workflow for randomized sampling. Behavior Multi-Modal Signals for Analyzing Pain Responses to Thermal and Electrical Stimuli Sascha Gruss1, Mattis Geiger2, Philipp Werner3, Oliver Wilhelm2, Harald C. Traue1, Ayoub Al-Hamadi3, Steffen Walter1 1Department of Medical Psychology, Ulm University, 2Department of Individual Differences and Psychological Assessment, Ulm University, 3Department of Neuro-Information Technology, University of Magdeburg This article focuses on the experimental elicitation of pain through heat (thermal) and electrical stimulation while recording physiological, visual, and paralinguistic responses. It aims at collecting valid multimodal data for analyzing pain based on its intensity, quality, and duration. Immunology and Infection Analysis of Shear Flow-induced Migration of Murine Marginal Zone B Cells In Vitro Kerry Tedford1, Laura Tech1, Michael Steiner1, Mark Korthals1, Klaus-Dieter Fischer1 1Institute of Biochemistry and Cell Biology, OVGU University of Magdeburg Marginal zone B cells (MZBs) respond to the force of shear flow by re-orienting their migration path up the flow. This protocol shows how to record and analyze the migration using a fluidics unit, pump, microscope imaging system, and free software. Developmental Biology High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo Nicole Meyer1, Thomas Schüler2, Ana Claudia Zenclussen1 1Experimental Obstetrics and Gynecology, Medical Faculty, Otto-von-Guericke University, 2Institute of Molecular and Clinical Immunology, Medical Faculty, Otto-von-Guericke University Here we describe the technique of high frequency ultrasound for in vivo analysis of fetuses in mice. This method allows the follow-up of fetuses and the analysis of placental parameters as well as maternal and fetal blood flow throughout pregnancy. Medicine Intravital Microscopy of Monocyte Homing and Tumor-Related Angiogenesis in a Murine Model of Peripheral Arterial Disease Martin Wagner1, Claudia Baer1, Werner Zuschratter2, Monika Riek-Burchardt3, Christian Deffge1, Soenke Weinert1, Jerry C Lee4, Ruediger C Braun-Dullaeus1, Joerg Herold1 1Department of Cardiology and Angiology, University of Magdeburg, 2Leibniz Institute for Neurobiology, 3Institute of Molecular and Clinical Immunology, University of Magdeburg, 4 Monocytes are important mediators of arteriogenesis in the context of peripheral arterial disease. Using a basement membrane-like matrix and intravital microscopy, this protocol investigates monocyte homing and tumor-related angiogenesis after monocyte injection in the femoral artery ligation murine model. Neuroscience High Resolution Quantitative Synaptic Proteome Profiling of Mouse Brain Regions After Auditory Discrimination Learning Angela Kolodziej1, Karl-Heinz Smalla1, Sandra Richter2, Alexander Engler1, Rainer Pielot1,3, Daniela C. Dieterich3, Wolfgang Tischmeyer1, Michael Naumann2, Thilo Kähne2 1Leibniz Institute for Neurobiology (LIN), 2Institute of Experimental Internal Medicine, Medical School, Otto von Guericke University Magdeburg, 3Institute of Pharmacology and Toxicology, Medical School, Otto von Guericke University The identification of molecules and pathways controlling synaptic plasticity and memory is still a major challenge in neuroscience. Here, a workflow is described addressing the relative quantification of synaptic proteins supposedly involved in the molecular reorganization of synapses during learning and memory consolidation in an auditory learning paradigm. Behavior Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning Max F.K. Happel1,2, Matthias Deliano*1, Frank W. Ohl*1,2,3 1Leibniz Institute for Neurobiology, Magdeburg, Germany, 2Otto-von-Guericke University, Magdeburg, Germany, 3Center for Behavioral Brain Sciences (CBBS), Magdeburg, Germany Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception. Immunology and Infection Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes Martin Wagner1, Helen Koester1, Christian Deffge1, Soenke Weinert1, Johannes Lauf1, Alexander Francke2, Jerry Lee3, R. C. Braun- Dullaeus1, Joerg Herold1 1Department for Cardiology, Angiology and Pneumology, Otto von Guericke University Magdeburg, 2Herzzentrum Dresden, Universitätsklinikum an der Technischen Universität Dresden, Technische Universität Dresden, 3Department of Public Health and Primary Care, University of Cambridge Here we present a protocol that generates large amounts of murine monocytes from heterogeneous bone marrow for translational applications. In comparison to others, this new method helps reduce the number of sacrificed animals and lowers costs by avoiding expensive methods such as high gradient magnetic cell separation (MACS). Immunology and Infection Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies Marcus Gereke1,2, Andrea Autengruber1,2, Lothar Gröbe3, Andreas Jeron2, Dunja Bruder1,2, Sabine Stegemann-Koniszewski1 1Research Group Immune Regulation, Helmholtz Centre for Infection Research, 2Research Group Infection Immunology, Institute of Medical Microbiology, Otto-von-Guericke University, 3Department of Experimental Immunology, Helmholtz Centre for Infection Research We describe the rapid isolation of primary murine type II alveolar epithelial cells (AECII) by flow cytometric negative selection. These AECII show high viability and purity and are suitable for a wide range of functional and molecular studies regarding their role in respiratory conditions such as autoimmune or infectious diseases. Immunology and Infection Direct Observation of Phagocytosis and NET-formation by Neutrophils in Infected Lungs using 2-photon Microscopy Mike Hasenberg1, Anja Köhler1, Susanne Bonifatius1, Andreas Jeron2, Matthias Gunzer1 1Institute for Molecular and Clinical Immunology, Otto-von-Guericke University Magdeburg, 2Department of Immunoregulation, Helmholtz Center for Infection Research We show, how to use 2-photon microscopy for the observation of the dynamics of neutrophil granulocytes in infected lungs while they phagocytose pathogens or produce neutrophil extracellular traps (NETs).