Institut du Cerveau et de la Moelle epiniere, ICM 4 articles published in JoVE Immunology and Infection Three-Dimensional Imaging of the Vertebral Lymphatic Vasculature and Drainage using iDISCO+ and Light Sheet Fluorescence Microscopy Laurent Jacob*1, Jose Brito*1,2, Jean-Leon Thomas1,3 1Université Pierre et Marie Curie Paris, Sorbonne Université, Institut du Cerveau et de la Moelle Epinière, 2Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, 3Department of Neurology, Yale University School of Medicine A protocol is presented combining tissue clearing with light sheet fluorescence microscopy (LSFM) to obtain three-dimensional and cellular resolution images of the lymphatic vessels and lymph nodes (LNs) collecting the cerebrospinal fluid (CSF) and spinal epidural fluid. Neuroscience Generation of Oligodendrocytes and Oligodendrocyte-Conditioned Medium for Co-Culture Experiments Elisa Mazuir1, Anne-Laure Dubessy1,2, Loane Wallon1,3, Marie-Stéphane Aigrot1, Catherine Lubetzki1,2, Nathalie Sol-Foulon1 1Sorbonne Université, Inserm, CNRS, UMR7225, Institut du Cerveau et de la Moelle épinière, ICM, 2Assistance Publique-Hôpitaux de Paris, GH Pitié Salpêtrière, 3Ecole Supérieure des Techniques de Biologie Appliquée Herein, we display an efficient method for the purification of oligodendrocytes and production of oligodendrocyte-conditioned medium that can be used for co-culture experiments. Immunology and Infection Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry Elodie Martin1,2,3, Mohamed El-Behi1,2,3, Bertrand Fontaine1,2,3,4, Cecile Delarasse1,2,3 1Inserm U 1227, CNRS UMR 7225, 2Sorbonne Universités, UPMC, University of Paris, 3Institut du Cerveau et de la Moelle épinière, ICM, 4AP-HP, Hôpital de la Pitié Salpêtrière; This protocol provides an analysis of the macrophage subpopulations in the adult mouse central nervous system by flow cytometry and is helpful for the study of multiple markers expressed by these cells. Neuroscience Induction of an Isoelectric Brain State to Investigate the Impact of Endogenous Synaptic Activity on Neuronal Excitability In Vivo Tristan Altwegg-Boussac1,2,3,4, Séverine Mahon1,2,3,4, Mario Chavez1,2,3,4, Stéphane Charpier1,2,3,4, Adrien E. Schramm1,2,3,4 1Inserm U1127, 2CNRS UMR 7225, 3UPMC Univ Paris 06, UMR S 1127, Sorbonne Universités, 4Institut du Cerveau et de la Moelle épinière (ICM) This procedure performs long-lasting in vivo intracellular recordings from single neurons during physiologically relevant cerebral states and after complete abolition of ongoing electrical activities, resulting in an isoelectric brain state. The physiological constants of the animal are carefully monitored during the transition to the artificial comatose condition.
