University of Wuerzburg View Institution's Website 8 articles published in JoVE Behavior A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test Marta Rattka1, Felix Fluri1, Miloš Krstić1, Esther Asan2, Jens Volkmann1 1Department of Neurology, University Hospital Wuerzburg, 2Institute of Anatomy and Cell Biology, University Wuerzburg Deep brain stimulation (DBS) is an effective treatment option for Parkinson's disease. We established a study design to screen novel stimulation paradigms in rats. The protocol describes the use of the staircase test and cylinder test for motor outcome assessment in DBS treated hemiparkinsonian rats. Bioengineering A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds Claudia Göttlich*1, Lena C. Müller*1, Meik Kunz*3, Franziska Schmitt1, Heike Walles1,4, Thorsten Walles2, Thomas Dandekar3, Gudrun Dandekar1,4, Sarah L. Nietzer1 1Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Wuerzburg, 2Department of Cardiothoracic Surgery, University Hospital Wuerzburg, 3Department of Bioinformatics, University Wuerzburg, 4Translational Center Wuerzburg, Fraunhofer Institute Interfacial Engineering and Biotechnology IGB We present a three-dimensional (3D) lung cancer model based on a biological collagen scaffold to study sensitivity towards non-small-cell-lung-cancer-(NSCLC)-targeted therapies. We demonstrate different read-out techniques to determine the proliferation index, apoptosis and epithelial-mesenchymal transition (EMT) status. Collected data are integrated into an in silico model for prediction of drug sensitivity. Biology Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED) Samira Samtleben1, Juliane Jaepel1,2, Caroline Fecher1, Thomas Andreska1, Markus Rehberg3, Robert Blum1 1Institute for Clinical Neurobiology, University of Wuerzburg, 2Department of Synapses - Circuits - Plasticity, Max Planck Institute of Neurobiology, Martinsried, 3Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians University of Munich Targeted-esterase induced dye loading (TED) supports the analysis of intracellular calcium store dynamics by fluorescence imaging. The method bases on targeting of a recombinant Carboxylesterase to the endoplasmic reticulum (ER), where it improves the local unmasking of synthetic low-affinity Ca2+ indicator dyes in the ER lumen. Neuroscience Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model Malgorzata Burek1, Ellaine Salvador1, Carola Y. Förster1 1Klinik und Poliklinik für Anästhesiologie, University of Wurzburg This method describes how to isolate and immortalize microvascular endothelial cells from mouse brain. We describe a step-by-step protocol starting from the homogenization of brain tissue, digestion steps, seeding and immortalization of the cells. Usually, it takes about five weeks to obtain a homogenous, immortalized microvascular endothelial cell line. 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. Biology The ITS2 Database Benjamin Merget1,2, Christian Koetschan1, Thomas Hackl1, Frank Förster1, Thomas Dandekar1, Tobias Müller1, Jörg Schultz1, Matthias Wolf1 1Department of Bioinformatics, Biocenter, University of Würzburg, 2Institute of Pharmacy and Food Chemistry, University of Würzburg The ITS2 Database is a workbench for phylogenetic inference simultaneously considering sequence and secondary structure of the internal transcribed spacer 2. This includes data collection with accurate annotation, structure prediction, multiple sequence-structure alignment and fast tree calculation. In a nutshell, this workbench simplifies first phylogenetic analyses to a few clicks. Immunology and Infection Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry Juan C. Garcia-Betancur1, Ana Yepes1, Johannes Schneider1, Daniel Lopez1 1Institute for Molecular Infection Biology (IMIB), University of Würzburg Microbial biofilms are generally constituted by distinct subpopulations of specialized cells. Single-cell analysis of these subpopulations requires the use of fluorescent reporters. Here we describe a protocol to visualize and monitor several subpopulationswithin B. subtilis biofilms using fluorescence microscopy and flow cytometry. 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.