University of Tübingen View Institution's Website 11 articles published in JoVE Immunology and Infection A Galleria mellonella Oral Administration Model to Study Commensal-Induced Innate Immune Responses Anna Lange1, Andrea Schäfer1, Julia-Stefanie Frick1 1Department for Medical Microbiology, Hygiene, Interfacultary Institute for Microbiology, Infection Medicine, University of Tübingen Here, we provide a detailed protocol for an oral administration model using Galleria mellonella larvae and how to characterize induced innate immune responses. Using this protocol, researchers without practical experience will be able to use the G. mellonella force-feeding method. Engineering 3D Printing of Biomolecular Models for Research and Pedagogy Eduardo Da Veiga Beltrame1, James Tyrwhitt-Drake2, Ian Roy3, Raed Shalaby4, Jakob Suckale4, Daniel Pomeranz Krummel5 1Department of Physics, Brandeis University, 2Bioinformatics and Computational Biosciences Branch (BCBB), NIH/NIAID/OD/OSMO/OCICB, 3Library/LTS/MakerLab, Brandeis University, 4Interfaculty Institute of Biochemistry (IFIB), University of Tübingen, 5Winship Cancer Institute, Emory University School of Medicine Physical models of biomolecules can facilitate an understanding of their structure-function for the researcher, aid in communication between researchers, and serve as an educational tool in pedagogical endeavors. Here, we provide detailed guidance for the 3D printing of accurate models of biomolecules using fused filament fabrication desktop 3D printers. Neuroscience Combined Optogenetic and Freeze-fracture Replica Immunolabeling to Examine Input-specific Arrangement of Glutamate Receptors in the Mouse Amygdala Sabine Schönherr1, Anna Seewald1, Yu Kasugai1, Daniel Bosch2, Ingrid Ehrlich2, Francesco Ferraguti1 1Department of Pharmacology, Medical University of Innsbruck, 2Hertie Institute for Clinical Brain Research and Centre for Integrative Neuroscience, University of Tübingen This article illustrates how the expression of neurotransmitter receptors can be quantified and the pattern analyzed at synapses with identified pre and postsynaptic elements using a combination of viral transduction of optogenetic tools and the freeze-fracture replica immunolabeling technique. Neuroscience Imaging Ca2+ Dynamics in Cone Photoreceptor Axon Terminals of the Mouse Retina Manoj Kulkarni1,2, Timm Schubert1,3, Tom Baden1,2,3, Bernd Wissinger4,5, Thomas Euler1,2,3, Francois Paquet-Durand1 1Institute for Ophthalmic Research, University of Tübingen, 2Graduate School of Cellular & Molecular Neuroscience, University of Tübingen, 3Bernstein Centre for Computational Neuroscience, University of Tübingen, 4Molecular Genetics Laboratory, University of Tübingen, 5Centre for Ophthalmology, University of Tübingen We describe a protocol to monitor Ca2+ dynamics in the axon terminals of cone photoreceptors using an ex-vivo slice preparation of the mouse retina. This protocol allows comprehensive studies of cone Ca2+ signaling in an important mammalian model system, the mouse. Neuroscience In ovo Expression of MicroRNA in Ventral Chick Midbrain Carola Huber1, A. Alwin Prem Anand1, Manfred Mauz1, Peter Künstle1, Wolfgang Hupp1, Bernhard Hirt1, Andrea Wizenmann1 1Department of Experimental Embryology, Institute of Anatomy, University of Tübingen Ectopic expression is one technique to elucidate the microRNAs role in brain development. However, targeting specific areas using in ovo electroporation is challenging. Here, we show an efficient way to selectively electroporate ventral and dorsal midbrain regions. Medicine The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease Lena F. Burbulla1,2, Rejko Krüger1,2 1German Center for Neurodegenerative Diseases, DZNE, 2Laboratory of Functional Neurogenomics, Department of Neurodegenerative Diseases, Hertie Institute for Clinical Brain Research, University of Tübingen Fibroblasts from patients carrying mutations in Parkinson's disease-causing genes represent an easily accessible ex vivo model to study disease-associated phenotypes. Live cell imaging gives the opportunity to study morphological and functional parameters in living cells. Here we describe the preparation of human fibroblasts and subsequent monitoring of mitochondrial phenotypes. Bioengineering Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy Miriam Votteler1,2, Daniel A. Carvajal Berrio2, Marieke Pudlas2,3, Heike Walles2,4, Katja Schenke-Layland1,2 1Department of Thoracic and Cardiovascular Surgery and Inter-University Centre for Medical Technology Stuttgart-Tübingen (IZST), Eberhard Karls University, Tübingen, 2Department of Cell and Tissue Engineering, Fraunhofer Institute of Interfacial Engineering and Biotechnology (IGB) Stuttgart, Germany, 3Department for Medical Interfacial Engineering (IGVT), University of Stuttgart, Germany, 4Institute of Tissue Engineering and Regenerative Medicine, Julius-Maximillians University, Würzburg, Germany Raman spectroscopy is a suitable technique for the non-contact, label-free analysis of living cells, tissue-engineered constructs and native tissues. Source-specific spectral fingerprints can be generated and analyzed using multivariate analysis. Neuroscience Chromatin Immunoprecipitation from Dorsal Root Ganglia Tissue following Axonal Injury Elisa Floriddia*1,2, Tuan Nguyen*1, Simone Di Giovanni1 1Laboratory for NeuroRegeneration and Repair, Department of Neurology, Hertie Institute for Clinical Brain Research, University of Tuebingen, 2Graduate School for Cellular and Molecular Neuroscience, University of Tuebingen We present a method for chromatin immunoprecipitation from dorsal root ganglia tissue following axonal injury. The approach can be used to identify specific transcription factor binding sites and epigenetic modification of histone and DNA important for the regeneration of injured axons in both the peripheral and central nervous system. Biology In vivo Imaging of Intact Drosophila Larvae at Sub-cellular Resolution Yao Zhang*1,2, Petra Füger*1, Shabab B. Hannan1,2, Jeannine V. Kern1, Bronwen Lasky1, Tobias M. Rasse1 1Junior Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen, 2Graduate School of Cellular and Molecular Neuroscience, University of Tübingen This protocol describes a reliable method for anesthetization and imaging of intact Drosophila melanogaster larvae. We have utilized the volatile anesthetic desflurane to allow for repetitive imaging at sub-cellular resolution and re-identification of structures for up to a few days1. Biology Right Hemihepatectomy by Suprahilar Intrahepatic Transection of the Right Hemipedicle using a Vascular Stapler Ingmar Königsrainer1, Silvio Nadalin1, Alfred Königsrainer1 1Department of General, Visceral, and Transplant Surgery, Tübingen University Hospital This video describes a right hemihepatectomy with intrahepatic transection of the right hemipedicle leaving the hepatoduodenal ligament completely untouched. Biology Non-invasive 3D-Visualization with Sub-micron Resolution Using Synchrotron-X-ray-tomography Michael Heethoff1, Lukas Helfen2, Peter Cloetens2 1Department of Evolutionary Biology of Invertebrates, University of Tubingen, 2European Synchrotron Radiation Facility We used synchrotron X-ray tomography at the European Synchrotron Radiation Facility (ESRF) to non-invasively produce 3D tomographic datasets with a pixel-resolution of 0.7µm. Using volume rendering software, this allows the reconstruction of internal structures in their natural state without the artefacts produced by histological sectioning.