Helmholtz Zentrum Munchen 8 articles published in JoVE Biochemistry Single-Copy Gene Locus Chromatin Purification in Saccharomyces cerevisiae Atiqa Sajid1, Stephan Hamperl1 1Institute of Epigenetics and Stem Cells, Helmholtz Zentrum München This protocol presents a locus-specific chromatin isolation method based on site-specific recombination to purify a single-copy gene locus of interest in its native chromatin context from budding yeast, Saccharomyces cerevisiae. Biology Visualizing Scar Development Using SCAD Assay - An Ex-situ Skin Scarring Assay Pushkar Ramesh1, Haifeng Ye1, Bikram Dasgupta1, Hans-Günther Machens2, Yuval Rinkevich1 1Institute of Regenerative Biology and Medicine, Helmholtz Zentrum München, 2School of Medicine, Klinikum Rechts der Isar, Department of Plastic and Hand Surgery, Technical University of Munich This protocol describes the generation of a skin-fascia explant termed "SCar like tissue in A Dish" or SCAD. This model allows unprecedented visualization of single fibroblasts during scar formation. Neuroscience Cryo-section Dissection of the Adult Subependymal Zone for Accurate and Deep Quantitative Proteome Analysis Christian Friess1, Magdalena Götz1,2,3, Jacob Kjell1,2,4 1Division of Physiological Genomics, Biomedical Center, Ludwig Maximilian University of Munich, 2Institute for Stem Cell Research, Helmholtz Zentrum München, 3SYNERGY, Excellence Cluster Systems Neurology, University of Munich, 4Department of Clinical Neuroscience, Karolinska Institutet Cryo-section-dissection allows fresh, frozen preparation of the largest neurogenic niche in the murine brain for deep quantitative proteome analysis. The method is precise, efficient, and causes minimal tissue perturbation. Therefore, it is ideally suited for studying the molecular microenvironment of this niche, as well as other organs, regions, and species. Chemistry Setup of Capillary Electrophoresis-Inductively Coupled Plasma Mass Spectrometry (CE-ICP-MS) for Quantification of Iron Redox Species (Fe(II), Fe(III)) Bernhard Michalke1, Desiree Willkommen1,4, Vivek Venkataramani2,3 1Research Unit Analytical BioGeoChemistry, Helmholz Zentrum München-German Research Center for Environmental Health, 2Department of Hematology and Medical Oncology, University Medical Center Göttingen (UMG), 3Institute of Pathology, University Medical Center Göttingen (UMG), 4RECIPE Chemicals + Instruments GmbH This iron redox speciation method is based on capillary electrophoresis-inductively coupled plasma mass spectrometry with sample stacking combined with short analysis in one run. The method quickly analyzes and provides low limits of quantification for iron redox species across a diverse range of tissues and biofluid samples. Developmental Biology Electroporation Method for In Vivo Delivery of Plasmid DNA in the Adult Zebrafish Telencephalon Tamara Durovic1,2, Jovica Ninkovic1,3,4 1Institute of Stem Cell Research, Helmholtz Zentrum München, 2Graduate School of Systemic Neurosciences, Ludwig Maximilian University, 3Department of Cell Biology and Anatomy, Biomedical Center, Ludwig Maximilian University, 4Munich Cluster for Systems Neurology (SyNergy) Presented here is an electroporation method for plasmid DNA delivery and ependymoglial cell labeling in the adult zebrafish telencephalon. This protocol is a quick and efficient method to visualize and trace individual ependymoglial cells and opens up new possibilities to apply electroporation to a broad range of genetic manipulations. Developmental Biology Partial Lobular Hepatectomy: A Surgical Model for Morphologic Liver Regeneration Jonathan M. Tsai1,2, Irving L Weissman1,2, Yuval Rinkevich3,4 1Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 2Department of Developmental Biology, Stanford University School of Medicine, 3Comprehensive Pneumology Center, Institute of Lung Biology and Disease, Helmholtz Zentrum München, 4German Center for Lung Research (DZL) Here, we present a new method for partial resection of the left hepatic lobe in neonatal (day 0) mice. This new protocol is suitable for studying acute liver injury and injury response in the neonatal setting. Cancer Research Hyperpolarized 13C Metabolic Magnetic Resonance Spectroscopy and Imaging Eugen Kubala*1,2,3, Kim A. Muñoz-Álvarez*1, Geoffrey Topping1, Christian Hundshammer1,2, Benedikt Feuerecker1, Pedro A. Gómez3,4, Giorgio Pariani1,5,6, Franz Schilling1, Steffen J. Glaser2, Rolf F. Schulte3, Marion I. Menzel3, Markus Schwaiger1 1Department of Nuclear Medicine, Klinikum rechts der Isar, Technische Universität München, 2Department of Chemistry, Technische Universität München, 3GE Global Research, 4Zentralinstitut für Medizintechnik der Technischen Universität München (IMETUM), Technische Universität München, 5Institute for Biological and Medical Imaging (IBMI), Helmholtz Zentrum München, 6IDG Institute of Developmental Genetics, Helmholtz Zentrum München Dynamic nuclear polarization with subsequent sample dissolution has enabled real-time studies of metabolism in biological systems. Hyperpolarized [1-13C]pyruvate was used to study lactate dehydrogenase activity in a prostate carcinoma cell line in vitro. Bioengineering Universal Hand-held Three-dimensional Optoacoustic Imaging Probe for Deep Tissue Human Angiography and Functional Preclinical Studies in Real Time Xosé Deán-Ben1, Thomas Felix Fehm1,2, Daniel Razansky1,2 1Institute for Biological and Medical Imaging (IBMI), Helmholtz Zentrum München, 2Faculty of Medicine, Technische Universität München We provide herein a detailed description of the experimental protocol for imaging with a newly developed hand-held optoacoustic (photoacoustic) system for three-dimensional functional and molecular imaging in real time. The demonstrated powerful performance and versatility may define new application areas of the optoacoustic technology in preclinical research and clinical practice.
