3 articles published in JoVE
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.
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.
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.