Heinrich-Heine-University 10 articles published in JoVE Genetics Application of DNA Fingerprinting using the D1S80 Locus in Lab Classes Meike Siebers*1, Agatha Walla*1,2, Thea Rütjes1, Moritz-Fabian Müller4, Maria von Korff1,2,3 1Institute of Plant Genetics, Heinrich-Heine-University, 2Cluster of Excellence on Plant Sciences "SMART Plants for Tomorrow's Needs", 3Max Planck Institute for Plant Breeding Research, 4IBG-1: Biotechnology, Forschungszentrum Jülich GmbH Here we describe a simple protocol to generate DNA fingerprinting profiles by amplifying the VNTR locus D1S80 from epithelial cell DNA. Neuroscience Generation of Human Brain Organoids for Mitochondrial Disease Modeling Stephanie Le1, Laura Petersilie2, Gizem Inak1,4, Carmen Menacho-Pando1, Karl W. Kafitz2, Agnieszka Rybak-Wolf3, Nikolaus Rajewsky3, Christine R. Rose2, Alessandro Prigione1,5 1Department of General Pediatrics, Neonatology and Pediatric Cardiology, Duesseldorf University Hospital, Medical Faculty, Heinrich Heine University, 2Institute of Neurobiology, Heinrich Heine University, 3Berlin Institute for Medical Systems Biology (BIMSB), Max Delbrueck Center for Molecular Medicine (MDC), 4Seaver Autism Center for Research and Treatment, Icahn School of Medicine at Mount Sinai, 5Max Delbrueck Center for Molecular Medicine (MDC) We describe a detailed protocol for the generation of human induced pluripotent stem cell-derived brain organoids and their use in modeling mitochondrial diseases. Biochemistry Imaging of Podocytic Proteins Nephrin, Actin, and Podocin with Expansion Microscopy Eva Königshausen*1, Christina Theresa Schmitz*1, Lars Christian Rump1, Lorenz Sellin1 1Department of Nephrology, Medical Faculty, Heinrich-Heine-University The presented method enables visualization of fluorescently labeled cellular proteins with expansion microscopy leading to a resolution of 70 nm on a conventional microscope. Medicine Highly Sensitive Measurement of Glomerular Permeability in Mice with Fluorescein Isothiocyanate-polysucrose 70 Eva Königshausen1, Sebastian A. Potthoff1, Magdalena Woznowski1, Johannes Stegbauer1, Lars C. Rump1, Lorenz Sellin1 1Department of Nephrology, Medical Faculty, Heinrich-Heine-University Here, we present a protocol to test glomerular permeability in mice using a highly sensitive, nonradioactive tracer. This method allows repetitive urine analyses with small urine volumes. Biochemistry Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins Eva Königshausen1, Sebastian A. Potthoff1, Raphael Haase1, Catherine Meyer-Schwesinger2, Ernest Kaufmann1, L. Christian Rump1, Johannes Stegbauer1, Lorenz Sellin1, Ivo Quack1, Magdalena Woznowski1 1Department of Nephrology, Medical Faculty, Heinrich-Heine-University, 2Institute of Cellular and Integrative Physiology, University Medical Center Hamburg-Eppendorf Here we present a protocol for murine in vivo labeling of glomerular cell surface proteins with biotin. This protocol contains information on how to perfuse mouse kidneys, isolate glomeruli, and perform endogenous immunoprecipitation of the protein of interest. Neuroscience Using Optical Coherence Tomography and Optokinetic Response As Structural and Functional Visual System Readouts in Mice and Rats Michael Dietrich1, Christina Hecker1, Alexander Hilla2, Andrés Cruz-Herranz3, Hans-Peter Hartung1, Dietmar Fischer2, Ari Green3, Philipp Albrecht1 1Department of Neurology, Heinrich-Heine-University Düsseldorf, 2Department of Cell Physiology, Faculty of Biology and Biotechnology, Ruhr-University Bochum, 3Division of Neuroinflammation and Glial Biology, Department of Neurology, University of California San Francisco A detailed protocol for the assessment of structural and visual readouts in rodents by optical coherence tomography and optokinetic response is presented. The results provide valuable insights for ophthalmologic as well as neurologic research. Biochemistry Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis Andreas Weber1, Julia Christin Wehmeyer1, Vera Schmidt1, Artur Lichtenberg1, Payam Akhyari1 1Department of Cardiovascular Surgery, Medical Faculty, Heinrich-Heine-University In this protocol, we describe the complete workflow for rapid isolation of extracellular vesicles from human whole blood and characterization of specific markers by fluorescence-based nanoparticle-tracking analysis. The presented results show a high level of reproducibility and can be adjusted to cell culture supernatants. Behavior Combined Invasive Subcortical and Non-invasive Surface Neurophysiological Recordings for the Assessment of Cognitive and Emotional Functions in Humans Carlos Trenado1, Saskia Elben2, David Petri2, Jan Hirschmann2, Stefan J. Groiss1,2, Jan Vesper3, Alfons Schnitzler1,2, Lars Wojtecki1,2 1Institute of Clinical Neuroscience and Medical Psychology, Medical Faculty, Heinrich-Heine-University, 2Department of Neurology, Center for Movement Disorders and Neuromodulation, University Clinic Düsseldorf, 3Department of Neurosurgery, Functional Neurosurgery and Stereotaxy, Center for Movement Disorders and Neuromodulation, University Clinic Düsseldorf The present protocol aims at assessing cognitive-emotional functions in the basal ganglia by simultaneous neurophysiological recording of local field potentials and non-invasive brain cortical activity (EEG). The procedure is exemplified by the use of paradigms involving speech stimuli with emotional connotation or the Flanker task involving cognitive control. Neuroscience Experimental Strategies to Bridge Large Tissue Gaps in the Injured Spinal Cord after Acute and Chronic Lesion Nicole Brazda*1, Veronica Estrada*1, Christian Voss2,3, Klaus Seide3, Hoc Khiem Trieu2, Hans Werner Müller1 1Molecular Neurobiology Laboratory, Department of Neurology, Heinrich-Heine-University Medical Center, 2Institute of Microsystems Technology, Hamburg University of Technology, 3Biomechanical Laboratory, BG Trauma Center Hamburg Severe spinal cord injuries often result in tissue defects. Two possibilities are described to successfully bridge such gaps to promote tissue adaptation, regenerative responses and functional improvement in rats via implantation of a mechanical microconnector system after acute injury and five weeks after complete spinal cord transection. Engineering Coherent anti-Stokes Raman Scattering (CARS) Microscopy Visualizes Pharmaceutical Tablets During Dissolution Andrew L. Fussell1, Peter Kleinebudde2, Jennifer Herek1, Clare J. Strachan3, Herman L. Offerhaus1 1Optical Sciences Group, MESA+ Institute, University of Twente, 2Institute of Pharmaceutics and Biopharmaceutics, Heinrich-Heine University, 3Division of Pharmaceutical Technology, Faculty of Pharmacy, University of Helsinki Coherent anti-Stokes Raman scattering (CARS) microscopy is combined with an intrinsic flow-through dissolution setup to allow in situ and real-time visualization of the surface of pharmaceutical tablets undergoing dissolution. Using this custom-built setup, it is possible to correlate CARS videos with drug dissolution profiles recorded using inline UV absorption spectroscopy.