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Systems Imaging of the Immune Synapse

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The Immune Synapse

Part of the book series: Methods in Molecular Biology ((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.

Rachel Ambler and Xiangtao Ruan are shared first authors.

Robert F. Murphy and Christoph Wülfing are shared senior authors.

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Acknowledgments

The original research upon which these protocols are based was supported in part by the National Institutes of Health grant P41 GM103712, by National Science Foundation grants MCB1121793 and MCB1121919, and by the European Research Council grant PCIG11-GA-2012-321554.

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Correspondence to Robert F. Murphy or Christoph Wülfing .

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Ambler, R., Ruan, X., Murphy, R.F., Wülfing, C. (2017). Systems Imaging of the Immune Synapse. In: Baldari, C., Dustin, M. (eds) The Immune Synapse. Methods in Molecular Biology, vol 1584. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6881-7_25

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  • DOI: https://doi.org/10.1007/978-1-4939-6881-7_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6879-4

  • Online ISBN: 978-1-4939-6881-7

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