Heidelberg University View Institution's Website 6 articles published in JoVE Biology A Modified Surgical Model of Hind Limb Ischemia in ApoE-/- Mice using a Miniature Incision Kaixuan Yan1,2, Jiaxing Zheng1,2, Frank G. Zöllner3,4, Kay Schwenke1, Prama Pallavi1,2, Michael Keese1,2 1Department of Surgery, Medical Faculty Manheim, Heidelberg University, 2European Center of Angioscience ECAS, Medical Faculty Manheim, Heidelberg University, 3Computer-Assisted Clinical Medicine, Mannheim Institute for Intelligent Systems in Medicine, Medical Faculty Mannheim, Heidelberg University, 4Cooperative Core Facility Animal Scanner ZI, Medical Faculty Mannheim, Heidelberg University This article demonstrates an efficient surgical approach to establish acute ischemia in mice with a small incision. This approach can be applied by most research groups without any laboratory upgrades. Biology A Simple Approach to Perform TEER Measurements Using a Self-Made Volt-Amperemeter with Programmable Output Frequency Marianne Theile1, Linus Wiora1, Dominik Russ1, Jonas Reuter1, Hiroshi Ishikawa2, Christian Schwerk3, Horst Schroten3, Stefan Mogk1 1Interfaculty Institute of Biochemistry, University of Tübingen, 2Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 3Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University Here, we demonstrate how to set up an inexpensive volt-amperemeter with programmable output frequency that can be used with commercially available chopstick electrodes for transepithelial/endothelial electrical resistance measurements. Cancer Research Paramyxoviruses for Tumor-targeted Immunomodulation: Design and Evaluation Ex Vivo Johannes P.W. Heidbuechel1,2, Christine E. Engeland1,3 1Department of Translational Oncology, German Cancer Research Center (DKFZ) and National Center for Tumor Diseases (NCT), 2Faculty of Biosciences, Heidelberg University, 3Department of Medical Oncology, NCT and Heidelberg University Hospital This protocol describes a detailed workflow for the generation and ex vivo characterization of oncolytic viruses for expression of immunomodulators, using measles viruses encoding bispecific T cell engagers as an example. Application and adaptation to other vector platforms and transgenes will accelerate the development of novel immunovirotherapeutics for clinical translation. Developmental Biology Multimodal Hierarchical Imaging of Serial Sections for Finding Specific Cellular Targets within Large Volumes Irene U. Wacker1,2, Lisa Veith3, Waldemar Spomer2,4, Andreas Hofmann2,4, Marlene Thaler5, Stefan Hillmer6, Ulrich Gengenbach2,4, Rasmus R. Schröder1,2,3 1Cryo Electron Microscopy, Centre for Advanced Materials, Universität Heidelberg, 2Heidelberg Karlsruhe Research Partnership (HEiKA), 3Cryo Electron Microscopy, BioQuant, Universitätsklinikum Heidelberg, 4Institute for Automation and Applied Computer Science, Karlsruhe Institute of Technology (KIT), 5Carl Zeiss Microscopy GmbH, 6Electron Microscopy Core Facility, Universität Heidelberg This protocol targets specific cells in tissue for imaging at nanoscale resolution using a scanning electron microscope (SEM). Large numbers of serial sections from resin-embedded biological material are first imaged in a light microscope to identify the target and then in a hierarchical manner in the SEM. Medicine A Choroid Plexus Epithelial Cell-based Model of the Human Blood-Cerebrospinal Fluid Barrier to Study Bacterial Infection from the Basolateral Side Stefanie Dinner1, Julia Borkowski1, Carolin Stump-Guthier1, Hiroshi Ishikawa2, Tobias Tenenbaum1, Horst Schroten1, Christian Schwerk1 1Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 2Department of NDU Life Sciences, Nippon Dental University The epithelial cells of the choroid plexus (CP) form the blood-cerebrospinal fluid barrier (BCSFB). An in vitro model of the BCSFB employs human choroid plexus papilloma (HIBCPP) cells. This article describes culturing and basolateral infection of HIBCPP cells using a cell culture filter insert system. Immunology and Infection Engineering and Evolution of Synthetic Adeno-Associated Virus (AAV) Gene Therapy Vectors via DNA Family Shuffling Eike Kienle*1, Elena Senís*1, Kathleen Börner2, Dominik Niopek1, Ellen Wiedtke1, Stefanie Grosse1, Dirk Grimm1 1Cluster of Excellence CellNetworks, Department of Infectious Diseases, Virology, Heidelberg University, 2Department of Infectious Diseases, Virology, Heidelberg University We demonstrate the basic technique to molecularly engineer and evolve synthetic Adeno-associated viral (AAV) gene therapy vectors via DNA family shuffling. Moreover, we provide general guidelines and representative examples for selection and analysis of individual chimeric capsids with enhanced properties on target cells in culture or in mice.