Systems Imaging of the Immune Synapse

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1584)

Abstract

Three-dimensional live cell imaging of the interaction of T cells with antigen-presenting cells (APCs) visualizes the subcellular distributions of signaling intermediates during T cell activation at thousands of resolved positions within a cell. These information-rich maps of local protein concentrations are a valuable resource in understanding T cell signaling. Here, we describe a protocol for the efficient acquisition of such imaging data and their computational processing to create four-dimensional maps of local concentrations. This protocol allows quantitative analysis of T cell signaling as it occurs inside live cells with resolution in time and space across thousands of cells.

Key words

Live cell imaging Computational image analysis Spatiotemporal distributions Immunological synapse T cell activation 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.School of Cellular and Molecular MedicineUniversity of BristolBristolUK
  2. 2.Computational Biology Department, School of Computer ScienceCarnegie Mellon UniversityPittsburghUSA
  3. 3.Department of Biological SciencesCarnegie Mellon UniversityPittsburghUSA
  4. 4.Department of Biomedical EngineeringCarnegie Mellon UniversityPittsburghUSA
  5. 5.Department of Machine LearningCarnegie Mellon UniversityPittsburghUSA

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