Otto-von-Guericke University Magdeburg

14 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. Lavrik^{1 1}Translational 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 Volckmar^1,2, Laura Knop^1,2,3, Tatjana Hirsch¹, Sarah Frentzel^1,2, Christian Erck⁴, Marco van Ham⁴, Sabine Stegemann-Koniszewski*^1,2,5, Dunja Bruder*^{1,2 1}Immune 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.

• 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 Veluswamy^{1 1}Department 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 Sundmacher^1,2, Tanja Vidaković-Koch^{1 1}Max 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 Haybäck^4,5, Florian Kleinegger⁴, Anna Birkl-Töglhofer⁴, Stefanie Krassnig⁴, Christina Wodlej⁴, Peter Holzer², Daniel Kummer¹, Elisabeth Bock¹, Gerd Leitinger^{1 1}Department 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 Walter^{1 1}Department 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 Fischer^{1 1}Institute 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 Zenclussen^{1 1}Experimental 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 Herold^{1 1}Department 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 Pielot^1,3, Daniela C. Dieterich³, Wolfgang Tischmeyer¹, Michael Naumann², Thilo Kähne^{2 1}Leibniz 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. Happel^1,2, Matthias Deliano*¹, Frank W. Ohl*^{1,2,3 1}Leibniz 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 Herold^{1 1}Department 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 Gereke^1,2, Andrea Autengruber^1,2, Lothar Gröbe³, Andreas Jeron², Dunja Bruder^1,2, Sabine Stegemann-Koniszewski^{1 1}Research 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 Gunzer^{1 1}Institute 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).