Ruhr-University Bochum View Institution's Website 20 articles published in JoVE Neuroscience Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules Maximilian Wulf1,2, Katalin Barkovits-Boeddinghaus1,2, Paula Sommer1,2, Karin Schork1,2, Martin Eisenacher1,2, Peter Riederer3,4, Manfred Gerlach5, Steffen Kösters1,2, Britta Eggers*1,2, Katrin Marcus*1,2 1Medizinisches Proteom-Center, Medical Faculty, Ruhr-University Bochum, 2Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, 3Center of Mental Health; Clinic and Policlinic for Psychiatry, Psychosomatics and Psychotherapy, University Hospital Wuerzburg, 4Psychiatry Department of Clinical Research, University of Southern Denmark Odense University Hospital, 5Center of Mental Health, Department of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital of Wuerzburg, University of Wuerzburg A robust protocol is presented here for isolating neuromelanin granules from human post-mortem substantia nigra pars compacta tissue via laser microdissection. This revised and optimized protocol massively minimizes the required time for sample collection, reduces the required sample amount, and enhances the identification and quantification of proteins by LC-MS/MS analysis. Medicine Nerve Ultrasound Protocol to Detect Dysimmune Neuropathies Anna Lena Fisse1,2, Kalliopi Pitarokoili1,2, Ralf Gold1,2 1Department of Neurology, St. Josef-Hospital, Ruhr-University Bochum, 2Immunmediated Neuropathies Biobank (INHIBIT), Ruhr-University Bochum This article presents a protocol for nerve ultrasound in polyneuropathies to aid the diagnosis of inflammatory neuropathies. Engineering Treating Surfaces with a Cold Atmospheric Pressure Plasma using the COST-Jet Judith Golda1, Kerstin Sgonina1, Julian Held2, Jan Benedikt1, Volker Schulz-von der Gathen2 1Experimental Plasma Physics, Kiel University, 2Experimental Physics II, Ruhr-University Bochum This protocol is presented to characterize the setup, handling, and application of the COST-Jet for the treatment of diverse surfaces such as solids and liquids. Behavior Optogenetic Manipulation of Neuronal Activity to Modulate Behavior in Freely Moving Mice Laura Berg1,2, Jill Gerdey1,2, Olivia A. Masseck1,2 1Biology and Chemistry, Synthetic Biology, University of Bremen, 2Advanced Fluorescence Microscopy, Faculty of Biology and Biotechnology, Ruhr-Universität Bochum With optogenetic manipulation of specific neuronal populations or brain regions, behavior can be modified with high temporal and spatial resolution in freely moving animals. By using different optogenetic tools in combination with chronically implanted optical fibers, a variety of neuronal modulations and behavioral testing can be performed. Biology Collection of Skeletal Muscle Biopsies from the Superior Compartment of Human Musculus Tibialis Anterior for Mechanical Evaluation Anthony L. Hessel1,4, Daniel Hahn1,2, Markus de Marées3 1Faculty of Sport Science, Human Movement Science, Ruhr University Bochum, 2School of Human Movement and Nutrition Sciences, The University of Queensland, 3Faculty of Sport Science, Sports Medicine and Sports Nutrition, Ruhr University Bochum, 4Institute of Physiology II, University of Muenster This technical report describes a variation of the modified Bergström technique for the biopsy of the musculus tibialis anterior that limits fiber damage. Immunology and Infection A Cell Culture Model for Producing High Titer Hepatitis E Virus Stocks Toni Luise Meister*1, Mara Klöhn*1, Eike Steinmann1, Daniel Todt1 1Molecular and Medical Virology, Ruhr University Bochum Described here is an effective method on how to produce high viral titer stocks of hepatitis E virus (HEV) to efficiently infect hepatoma cells. With the presented method, both non-enveloped, as well as enveloped viral particles can be harvested and used for inoculating various cell lines. Medicine Patient Directed Recording of a Bipolar Three-Lead Electrocardiogram using a Smartwatch with ECG Function Alexander Samol1, Kristina Bischof1, Blerim Luani1, Dan Pascut1, Marcus Wiemer1, Sven Kaese1 1Department of Cardiology and Critical Care Medicine, Johannes Wesling University Hospital, Ruhr University Bochum We describe a protocol for the patient-directed registration of a three lead bipolar electrocardiogram by a smartwatch that functions identically to the Einthoven leads from standard electrocardiograms. This enables patients to record electrocardiograms on their own immediately after the onset of symptoms. Medicine Modified Single-Loop Reconstruction for Pancreaticoduodenectomy Andreas M. Luu1, Ilgar Aghalarov1, Chris Braumann1, Waldemar Uhl1, Orlin Belyaev1 1Department of General and Visceral Surgery, St. Josef Hospital, Ruhr University Bochum The technique of modified single-loop reconstruction following pancreaticoduodenectomy separates pancreatic secretion from bile, thus reducing the severity of POPF without prolonging the duration of surgery. Developmental Biology Assessment of Ultrastructural Neuroplasticity Parameters After In Utero Transduction of the Developing Mouse Brain and Spinal Cord David Lutz*1, Monika von Düring*1, Franco Corvace1, Luzie Augustinowski1, Anne-Kathrin Trampe1, Marzena Nowak2, Eckart Förster1 1Department of Neuroanatomy and Molecular Brain Research, Ruhr University Bochum, 2Central Animal Facility of the Medical Faculty, Ruhr University Bochum The combination of transmission electron microscopy and in utero transduction is a powerful approach for studying morphological changes in the fine ultrastructure of the nervous system during development. This combined method allows deep insights into changes in structural details underlying neuroplasticity with respect to their topographical representation. 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. Biology Investigating the Detrimental Effects of Low Pressure Plasma Sterilization on the Survival of Bacillus subtilis Spores Using Live Cell Microscopy Felix M. Fuchs1, Marina Raguse1,2,3, Marcel Fiebrandt2, Kazimierz Madela4, Peter Awakowicz2, Michael Laue4, Katharina Stapelmann3, Ralf Moeller1 1Department of Radiation Biology, Institute of Aerospace Medicine, Space Microbiology Research Group, German Aerospace Center (DLR e.V.), 2Institute of Electrical Engineering and Plasma Technology, Faculty of Electrical Engineering and Information Technology, Ruhr-University Bochum, 3Institute of Electrical Engineering and Plasma Technology, Faculty of Electrical Engineering and Information Technology, Biomedical Applications of Plasma Technology, Ruhr-University Bochum, 4Advanced Light and Electron Microscopy (ZBS 4), Robert Koch Institute This protocol illustrates the important consecutive steps required to assess the relevance of monitoring vitality parameter and DNA repair processes in reviving Bacillus subtilis spores after treatment with low pressure plasma by tracking fluorescence-labelled DNA repair proteins via time-resolved confocal microscopy and scanning electron microscopy. Chemistry Simple Methods for the Preparation of Non-noble Metal Bulk-electrodes for Electrocatalytic Applications Kai junge Puring1,2, Stefan Piontek1, Mathias Smialkowski1, Jens Burfeind2, Stefan Kaluza2, Christian Doetsch2, Ulf-Peter Apfel1 1Faculty of Chemistry and Biochemistry, Inorganic Chemistry I, Ruhr-University Bochum, 2Fraunhofer Institute for Environmental, Safety, and Energy Technology, UMSICHT A facile preparation method of electrodes using the bulk material Fe4.5Ni4.5S8 is presented. This method provides an alternative technique to conventional electrode fabrication and describes prerequisites for unconventional electrode materials including a straightforward electrocatalytic testing method. Medicine In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging Angelika Hoffmann1,2, Xavier Helluy2,3, Manuel Fischer2, Ann-Kristin Mueller4,5, Sabine Heiland2, Mirko Pham1,6, Martin Bendszus1, Johannes Pfeil4,5,7 1Department of Neuroradiology, Heidelberg University Hospital, 2Division of Experimental Radiology, Department of Neuroradiology, Heidelberg University Hospital, 3NeuroImaging Centre Research, Department of Neuroscience, Ruhr-University Bochum, 4Centre for Infectious Diseases, Parasitology Unit, Heidelberg University Hospital, 5German Centre for Infection Research (DZIF), 6Department of Neuroradiology, University of Würzburg, 7Center for Childhood and Adolescent Medicine, General Pediatrics, Heidelberg University Hospital We describe a mouse model of experimental cerebral malaria and show how inflammatory and microvascular pathology can be tracked in vivo using magnetic resonance imaging. Medicine Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry (LA-ICP-MS) Dominic J. Hare1,2, Kai Kysenius3, Bence Paul4, Beate Knauer5,6, Robert W. Hutchinson7, Ciaran O'Connor7, Fred Fryer8, Tom P. Hennessey8, Ashley I. Bush2, Peter J. Crouch3, Philip A. Doble1 1Elemental Bio-imaging Facility, University of Technology Sydney, 2Florey Institute of Neuroscience and Mental Health, The University of Melbourne, 3Department of Pathology, The University of Melbourne, 4School of Earth Sciences, The University of Melbourne, 5Research School, Ruhr University, 6Department of Physiology, Monash University, 7ESI Ltd., Bozeman, 8Agilent Technologies, Mulgrave Quantitatively mapping metals in tissue by laser ablation - inductively coupled plasma - mass spectrometry (LA-ICP-MS) is a sensitive analytical technique that can provide new insight into how metals participate in normal function and disease processes. Here, we describe a protocol for quantitatively imaging metals in thin sections of mouse neurological tissue. Neuroscience The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions Christine Gottschling1, Egor Dzyubenko1, Maren Geissler1, Andreas Faissner1 1Department of Cell Morphology and Molecular Neurobiology, Faculty of Biology and Biotechnology, Ruhr-University This protocol describes the indirect neuron-astrocyte coculture for compartmentalized analysis of neuron-glia interactions. Chemistry Light-driven Enzymatic Decarboxylation Katharina Köninger1, Marius Grote1, Ioannis Zachos1, Frank Hollmann2, Robert Kourist1 1Faculty of Biology and Biotechnology, Ruhr Universität Bochum, 2Biology and Biotechnology, Delft University of Technology We describe a protocol for the light-catalyzed generation of hydrogen peroxide — a cofactor for oxidative transformations. Engineering Experimental Methods for Investigation of Shape Memory Based Elastocaloric Cooling Processes and Model Validation Marvin Schmidt1,2, Johannes Ullrich2, André Wieczorek3, Jan Frenzel3, Gunther Eggeler3, Andreas Schütze1, Stefan Seelecke2 1Lab for Measurement Technology, Saarland University, 2Intelligent Material Systems Lab, Saarland University, 3Lab for Material Science, Ruhr Universität Bochum Experimental methods for investigation of solid state cooling processes and characterization of elastocaloric material properties of Shape Memory Alloys (SMA) are presented. A custom-built test rig has been designed for controlling and comprehensive monitoring of elastocaloric cooling processes. Furthermore, it provides a validation platform for thermomechanically coupled modeling approaches. Biology Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli Louise E. Bird1, Heather Rada1, Anil Verma1, Raphael Gasper2,3, James Birch4, Matthew Jennions4, Jan Lӧwe3, Isabel Moraes4, Raymond J. Owens1 1Oxford Protein Production Facility, Research Complex at Harwell, 2Protein Crystallography Group, Ruhr University, 3MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, 4Membrane Protein Laboratory, Diamond Light Source A streamlined approach to screening for the expression of recombinant membrane proteins in Escherichia coli based on fusion to green fluorescent protein is presented. Neuroscience Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning Sarah Starosta*1, Maik C. Stüttgen*1, Onur Güntürkün1 1Faculty of Psychology, Department of Biopsychology, Ruhr-University Bochum Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session. Neuroscience Lectin-based Isolation and Culture of Mouse Embryonic Motoneurons Rebecca Conrad1, Sibylle Jablonka2, Teresa Sczepan1, Michael Sendtner2, Stefan Wiese1, Alice Klausmeyer1 1Institute for Cellmorphology and molecular Neurobiology, Group for Cellbiology, Ruhr-University Bochum, 2Institute for Clinical Neurobiology, University of Wuerzburg An alternative way of isolating mouse embryonic motoneurons from the spinal cord is described. The method takes into account the fact that lectin can bind to the low affinity nerve growth factor receptor p75NTR. This lectin-based preplating allows a purification similar to that with a specific antibody against the p75NTR.
