Institut de la Vision 5 articles published in JoVE Neuroscience Cone-Enriched Cultures from the Retina of Chicken Embryos to Study Rod to Cone Cellular Interactions Géraldine Millet-Puel1, Myriam Pinault1, Marie Cordonnier1, Valérie Fontaine1, José-Alain Sahel1, Thierry Léveillard1 1Department of Genetics - Sorbonne Université, INSERM, CNRS, Institut de la Vision We describe a method to obtain primary cultures of cone photoreceptors from the retina of chicken embryos and its use for high content screening. Neuroscience In Utero Electroporation of Multiaddressable Genome-Integrating Color (MAGIC) Markers to Individualize Cortical Mouse Astrocytes Laura Dumas*1, Solène Clavreul*2, Jason Durand1, Edwin Hernandez-Garzon3, Lamiae Abdeladim4, Raphaëlle Barry-Martinet2, Alicia Caballero-Megido1, Emmanuel Beaurepaire4, Gilles Bonvento3, Jean Livet2, Karine Loulier1 1Université de Montpellier, INSERM, Institut des Neurosciences de Montpellier, 2Sorbonne Université, INSERM, CNRS, Institut de la Vision, 3Université Paris-Saclay, CEA, CNRS, MIRCen, Laboratoire des Maladies Neurodégénératives, 4Laboratory for Optics and Biosciences, Ecole polytechnique, CNRS, INSERM, IP Paris Astrocytes tile the cerebral cortex uniformly, making the analysis of their complex morphology challenging at the cellular level. The protocol provided here uses multicolor labeling based on in utero electroporation to single out cortical astrocytes and analyze their volume and morphology with a user-friendly image analysis pipeline. Medicine Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis Nacim Bouheraoua*1,2,3,4, Lea Jouve*1,2,3,4, Vincent Borderie1,2,3,4, Laurent Laroche1,2,3,4 1Quinze-Vingts National Ophthalmology Hospital, 2INSERM UMR S 968, Institut de la Vision, 3Sorbonne Universités, UPMC Univ Paris 06, 4CNRS, UMR 7210 Corneal collagen cross-linking (CXL) is the only conservative treatment currently available to halt keratoconus progression by improving the biomechanical rigidity of the corneal stroma. The aim of this manuscript is to highlight the methods of three different protocols of CXL: conventional CXL (C-CXL), accelerated CXL (A-CXL), and iontophoresis CXL (I-CXL). Neuroscience Vibratome Sectioning Mouse Retina to Prepare Photoreceptor Cultures Emmanuelle Clérin1,4,5,6, Ying Yang1,4,5,6, Valérie Forster2,4,5,6, Valérie Fontaine3,4,5,6, José-Alain Sahel4,5,6, Thierry Léveillard1,4,5,6 1Department of Genetics, UMR_S 968, Institut de la Vision, 2Department of Visual Information, UMR_S 968, Institut de la Vision, 3Exploratory Team, UMR_S 968, Institut de la Vision, 4Sorbonne Universités, Paris 06, UMR_S 968, Institut de la Vision, 5INSERM, U968, Institut de la Vision, 6CNRS, UMR_7210, Institut de la Vision Neural retina of a mouse aged 8 days is on top of a 4% gelatin block. After isolation of the photoreceptor layer (200 µm) by vibratome, the photoreceptors are seeded after mechanical and enzymatic dissociation for culture. The photoreceptor layer can be used for molecular, biochemical analyses or transplantation. Immunology and Infection Large-Scale Purification of Porcine or Bovine Photoreceptor Outer Segments for Phagocytosis Assays on Retinal Pigment Epithelial Cells Célia Parinot1,2,3, Quentin Rieu1,2,3, Jonathan Chatagnon1,2,3, Silvia C. Finnemann4, Emeline F. Nandrot1,2,3 1INSERM, U968, 2Sorbonne Universités, UPMC Paris 06, UMR_S 968, Institut de la Vision, 3CNRS, UMR_7210, 4Department of Biological Sciences, Center for Cancer, Genetic Diseases and Gene Regulation, Fordham University This article describes the protocol for the purification of photoreceptor outer segment fragments (POS) via ultracentrifugation from porcine/bovine retinae using homogenization and sucrose gradient centrifugation. This protocol allows the preparation of large stocks of POS aliquots, labeled or unlabeled, that can then be stored at -80 °C.
