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An Endothelial Planar Cell Model for Imaging Immunological Synapse Dynamics
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Studying the T Cell-Astrocyte Immune Synapse

George P Cribaro1, Elena Saavedra-López1, Paola V Casanova1

  • 1Neuroimmunity Research Group, Institut de Neurociències, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|March 4, 2017
PubMed

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).

Keywords:
3D reconstructionAstrocyteConfocal microscopyImmunofluorescenceT cellTissuec-SMACp-SMAC

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