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The Immune Synapse pp 383–398Cite as

Imaging Asymmetric T Cell Division

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

Abstract

Asymmetric cell division (ACD) controls cell fate decisions in model organisms such as Drosophila and C. elegans and has recently emerged as a mediator of T cell fate and hematopoiesis. The most appropriate methods for assessing ACD in T cells are still evolving. Here we describe the methods currently applied to monitor and measure ACD of developing and activated T cells. We provide an overview of approaches for capturing cells in the process of cytokinesis in vivo, ex vivo, or during in vitro culture. We provide methods for in vitro fixed immunofluorescent staining and for time-lapse analysis. We provide an overview of the different approaches for quantification of ACD of lymphocytes, discuss the pitfalls and concerns in interpretation of these analyses, and provide detailed methods for the quantification of ACD in our group.

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Acknowledgments

We would like to acknowledge members of the Russell lab (past and present) for the development of the methods described here.

Author information

Authors and Affiliations

  1. Faculty of Science, Engineering and Technology, Centre for Micro-Photonics, Swinburne University of Technology, Mail No H74, PO Box 218, Hawthorn, VIC, 3122, Australia

    Mirren Charnley & Sarah M. Russell

  2. Immune Signalling Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC, 3002, Australia

    Mirren Charnley & Sarah M. Russell

  3. Faculty of Science, Engineering and Technology, Biointerface Engineering, Swinburne University of Technology, Hawthorn, VIC, 3122, Australia

    Mirren Charnley

  4. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia

    Sarah M. Russell

  5. Department of Pathology, The University of Melbourne, Parkville, VIC, 3052, Australia

    Sarah M. Russell

Authors
  1. Mirren Charnley
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  2. Sarah M. Russell
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Corresponding author

Correspondence to Sarah M. Russell .

Editor information

Editors and Affiliations

  1. Department of Life sciences, University of Siena, Siena, Siena, Italy

    Cosima T. Baldari

  2. University of Oxford, Kennedy Institute of Rheumatology, Headington, Oxford, United Kingdom

    Michael L. Dustin

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Charnley, M., Russell, S.M. (2017). Imaging Asymmetric T Cell Division. 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_23

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  • DOI: https://doi.org/10.1007/978-1-4939-6881-7_23

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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

  • eBook Packages: Springer Protocols

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