University of Tubingen 10 articles published in JoVE Immunology and Infection Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test Julia Häring*1, Tanja Michel*1, Matthias Becker1, Daniel Junker1, Tatia Tchitchagua2, Olaf Leschnik2, Berit Lange3,4, Stefanie Castell3,4, Gérard Krause3,4, Monika Strengert3, Alex Dulovic1, Nicole Schneiderhan-Marra1 1NMI Natural and Medical Sciences Institute at the University of Tübingen, 2Department of Neurology, Sächsisches Krankenhaus Rodewisch, 3Department of Epidemiology, Helmholtz Centre for Infection Research, 4German Centre for Infection Research (DZIF) A three-channel dual-reporter fluorescence flow analysis system was used to develop a bead-based multiplex immunoassay that simultaneously evaluates serum samples for IgG and IgM elicited against multiple antigens of different Borrelia species that cause Lyme borreliosis in Europe and North America. Medicine Laser-Induced Action Potential-Like Measurements of Cardiomyocytes on Microelectrode Arrays for Increased Predictivity of Safety Pharmacology Jasmin Schaefer1, Timm Danker1, Karin Gebhardt1, Udo Kraushaar1 1NMI Natural and Medical Sciences Institute at the University of Tübingen, Germany, The combination of laser poration and microelectrode arrays (MEA) allows action potential-like recordings of cultivated primary and stem cell-derived cardiomyocytes. The waveform shape provides superior insight into test compounds' mode of action than standard recordings. It links patch-clamp and MEA readout to further optimize cardio safety research in the future. Biology Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to Plasmodiophora brassicae Deeksha Singh1, Sara Blicharz1, Karolina Stefanowicz1, Laura Ragni2, Kornel Michalak3, Agnieszka Bagniewska-Zadworna3, Robert Malinowski1 1Integrative Plant Biology Team, Institute of Plant Genetics Polish Academy of Sciences, 2ZMBP-Center for Plant Molecular Biology, University of Tübingen, 3Department of General Botany, Institute of Experimental Biology, Faculty of Biology, Adam Mickiewicz University The present protocol describes an optimized method for the histological observation of galls induced by Plasmodiophora brassicae. Vibratome sections of hypocotyls are cleared before fluorescence imaging to study the involvement of transcription factors and phytohormones during disease progression. This protocol overcomes resin embedding limitations, enabling in planta visualization of fluorescent proteins. Biology Optical Sectioning and Visualization of the Intervertebral Disc from Embryonic Development to Degeneration Florian Christof Bonnaire1,2, Martina Feierabend3, Julius Michael Wolfgart1,4, Wolfram Breuer5, Christian Walter2, Ulf Krister Hofmann1,2, Marina Danalache1 1Laboratory of Cell Biology, Department of Orthopedic Surgery, University Hospital of Tübingen, 2Department of Orthopedic Surgery, University Hospital of Tübingen, 3Institute for Bioinformatics and Medical Informatics, Faculty of science of the University of Tübingen, 4Medical faculty of the University of Tübingen, 5Bavarian Health and Food Safety Authority We present a method to investigate spatial chondrocyte organization in the anulus fibrosus of the intervertebral disc using an optical sectioning method. Neuroscience Long-Term, Serum-Free Cultivation of Organotypic Mouse Retina Explants with Intact Retinal Pigment Epithelium Soumaya Belhadj*1, Arianna Tolone*1, Gustav Christensen1, Soumyaparna Das1, Yiyi Chen1, François Paquet-Durand1 1Institute for Ophthalmic Research, University of Tübingen The protocol describes organotypic explants of mouse neuroretina, cultivated together with its retinal pigment epithelium (RPE), in R16 defined medium, free of serum and antibiotics. This method is relatively simple to perform, less expensive, and time-consuming when compared to in vivo experiments, and can be adapted to numerous experimental applications. Biology Automated Production of Human Induced Pluripotent Stem Cell-Derived Cortical and Dopaminergic Neurons with Integrated Live-Cell Monitoring Ashutosh Dhingra*1, Joachim Täger*1, Elisangela Bressan*1, Salvador Rodriguez-Nieto1, Manmeet-Sakshi Bedi1, Stefanie Bröer1, Eldem Sadikoglou1, Noémia Fernandes2, Melissa Castillo-Lizardo1, Patrizia Rizzu2, Peter Heutink1,3 1Genome Biology of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), 2Applied Genomics for Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), 3Department for Neurodegenerative Diseases, Hertie Institute for Clinical Brain Research, University of Tübingen We show the automation of human induced pluripotent stem cell (hiPSC) cultures and neuronal differentiations compatible with automated imaging and analysis. Bioengineering Application of Atomic Force Microscopy to Detect Early Osteoarthritis Marina Danalache*1, Aadhya Tiwari*1, Viktor Sigwart1,2, Ulf Krister Hofmann1,3 1Laboratory of Cell Biology, Department of Orthopaedic Surgery, University Hospital of Tübingen, 2Medical faculty of the University of Tübingen, 3Department of Orthopaedic Surgery, University Hospital of Tübingen We present a method to investigate early osteoarthritic changes at the cellular level in articular cartilage by using atomic force microscopy (AFM). 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. Behavior Brain State-dependent Brain Stimulation with Real-time Electroencephalography-Triggered Transcranial Magnetic Stimulation Maria-Ioanna Stefanou1,2, David Baur1,2, Paolo Belardinelli1,2, Til Ole Bergmann1,2, Corinna Blum1,2, Pedro Caldana Gordon1,2, Jaakko O. Nieminen1,2,3, Brigitte Zrenner1,2, Ulf Ziemann1,2, Christoph Zrenner1,2 1Department of Neurology & Stroke, University of Tübingen, 2Hertie Institute for Clinical Brain Research, University of Tübingen, 3Department of Neuroscience and Biomedical Engineering, Aalto University This paper describes real-time electroencephalography-triggered transcranial magnetic stimulation to study and modulate human brain networks. Biology Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo Christopher F. Schuster1, Ralph Bertram1 1Department of Microbial Genetics, Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), Faculty of Science, University of Tübingen We here describe a fluorescence based primer extension method to determine transcriptional starting points from bacterial transcripts and RNA processing in vivo using an automated gel sequencer.