title: Studying the T Cell-Astrocyte Immune Synapse.
abstract: In this chapter, we describe the technical details to visualize and analyze effector immunological synapses between T cells and astrocytes in the brain with high-resolution confocal imaging. This procedure is critical for the optimal and even penetration of labeling antibodies within the nerve tissue to obtain accurate staining and allow a uniform three-dimensional analysis of the T cell-astrocyte interactions. We emphasize here the comprehensive exploration of the tissue and analysis with confocal microscope as well as the display of microanatomical details of the three-dimensional reconstruction for interface visualization (including peripheral and central supramolecular activation clusters, effector molecules, and other organelles such as microtubule organizing centers (MTOCs) and Golgi apparatus).
authors.affiliation: Neuroimmunity Research Group, Institut de Neurociències, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: George P
authors.initials: GP
authors.lastname: Cribaro
authors.affiliation: Neuroimmunity Research Group, Institut de Neurociències, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Saavedra-López
authors.affiliation: Neuroimmunity Research Group, Institut de Neurociències, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Paola V
authors.initials: PV
authors.lastname: Casanova
authors.affiliation: Neuroimmunity Research Group, Institut de Neurociències, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Rodríguez
authors.affiliation: Neuroimmunity Research Group, Institut de Neurociències, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain. carlos.barcia@uab.es.
authors.forename: Carlos
authors.initials: C
authors.lastname: Barcia
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000276:immunology*; D002450:Cell Communication / Q000276:immunology*; D006056:Golgi Apparatus / Q000276:immunology; D006801:Humans; D054992:Immunological Synapses / Q000276:immunology*; D018613:Microscopy, Confocal / Q000379:methods; D022101:Microtubule-Organizing Center / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
keywords: 3D reconstruction; Astrocyte; Confocal microscopy; Immunofluorescence; T cell; Tissue; c-SMAC; p-SMAC