Helmholtz Center Munich View Institution's Website 7 articles published in JoVE Biology Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics Mateusz Strzelecki1,4, Kelvin Yin1, Carlos Talavera-López2,3, Celia P. Martinez-Jimenez1,4 1Helmholtz Pioneer Campus (HPC), Helmholtz Munich, 2Institute of Computational Biology, Computational Health Department, Helmholtz Munich, 3Division of Infectious Diseases and Tropical Medicine, Ludwig-Maximillian-Universität Klinikum, 4TUM School of Medicine, Technical University of Munich Here, we present a protocol for isolating nuclei from flash-frozen, archived liver tissues for single-nucleus RNA-seq, ATAC-seq, and joint multiomics (RNA-seq and ATAC-seq). Neuroscience Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry Asa S. Hidmark1, Peter P. Nawroth1,2,3, Thomas Fleming1,3 1Department of Medicine I and Clinical Chemistry, University Hospital of Heidelberg, 2Institute for Diabetes and Cancer IDC Helmholtz Center Munich, Germany & Joint Heidelberg-IDC Translational Diabetes Program, 3German Center for Diabetes Research (DZD) Quantitative analysis of cell content within the murine sciatic nerve is difficult due to the scarcity of the tissue. This protocol describes a method for tissue digestion and preparation that provides sufficient cells for flow cytometry analysis of immune cell populations from nerves of individual mice. Immunology and Infection Adenoviral Transduction of Naive CD4 T Cells to Study Treg Differentiation Sebastian C. Warth1, Vigo Heissmeyer1 1Institute for Molecular Immunology, Helmholtz Zentrum München Adenoviral gene transfer into naive CD4 T cells with transgenic expression of the Coxsackie adenovirus receptor enables the molecular analysis of regulatory T cell differentiation in vitro. Medicine Time-lapse Imaging of Primary Preneoplastic Mammary Epithelial Cells Derived from Genetically Engineered Mouse Models of Breast Cancer Rebecca E. Nakles1, Sarah L. Millman1, M. Carla Cabrera1, Peter Johnson1,2, Susette Mueller1,2, Philipp S. Hoppe3, Timm Schroeder3, Priscilla A. Furth1,2,4,5 1Department of Oncology, Georgetown University, 2Lombardi Comprehensive Cancer Center, Georgetown University, 3Stem Cell Dynamics, Helmholtz Zentrum München - German Research Center for Environmental Health, 4Department of Medicine, Georgetown University, 5Department of Nanobiomedical Science and WCU Research Center of Nanobiomedical Science, Dankook University Time-lapse imaging is used to assess behavior of primary preneoplastic mammary epithelial cells derived from genetically engineered mouse models of breast cancer risk to determine if there are correlations between specific behavioral parameters and distinct genetic lesions. Bioengineering Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy Felix Spira1, Julia Dominguez-Escobar1, Nikola Müller1,2, Roland Wedlich-Söldner1 1AG Cellular Dynamics and Cell Patterning, Max Planck Institute of Biochemistry, 2Helmholtz Zentrum München Total Internal Reflection Fluorescence (TIRF) microscopy is a powerful approach to observe structures close to the cell surface at high contrast and temporal resolution. We demonstrate how TIRF can be employed to study protein dynamics at the cortex of cell wall-enclosed bacterial and fungal cells. Biology Mesoscopic Fluorescence Tomography for In-vivo Imaging of Developing Drosophila Claudio Vinegoni1, Daniel Razansky2, Chrysoula Pitsouli3, Norbert Perrimon3, Vasilis Ntziachristos2, Ralph Weissleder1 1Center for Systems Biology, Massachusetts General Hospital, 2Institute for Biological and Medical Imaging (IBMI), Technical University of Munich and Helmholtz Center Munich, 3Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute Mesoscopic fluorescence tomography operates beyond the penetration limits of tissue-sectioning fluorescence microscopy. The technique is based on multi-projection illumination and a photon transport description. We demonstrate in-vivo whole-body 3D visualization of the morphogenesis of GFP-expressing wing imaginal discs in Drosophila melanogaster. Biology Born Normalization for Fluorescence Optical Projection Tomography for Whole Heart Imaging Claudio Vinegoni1,2, Daniel Razansky3, Jose-Luiz Figueiredo1,2, Lyuba Fexon1,2, Misha Pivovarov1,2, Matthias Nahrendorf1,2, Vasilis Ntziachristos3, Ralph Weissleder1,2 1Center for Systems Biology, Harvard Medical School, 2Center for Systems Biology, MGH - Massachusetts General Hospital, 3Institute for Biological and Medical Imaging, Technical University of Munich and Helmholtz Center Munich We suggest a Born normalized approach for Optical Projection Tomography (BnOPT) that accounts for the absorption properties of imaged samples to obtain accurate and quantitative fluorescence tomographic reconstructions. We use the proposed algorithm to reconstruct the fluorescence molecular probe distribution within small animal organs.
