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Imaging Asymmetric T Cell Division

  • Mirren Charnley
  • Sarah M. RussellEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Asymmetric cell division T Cells Lymphocytes Cell polarity Fluorescence microscopy Image quantification Thymocyte Cell fate determinants 

Notes

Acknowledgments

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

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Faculty of Science, Engineering and Technology, Centre for Micro-PhotonicsSwinburne University of TechnologyHawthornAustralia
  2. 2.Immune Signalling LaboratoryPeter MacCallum Cancer CentreEast MelbourneAustralia
  3. 3.Faculty of Science, Engineering and Technology, Biointerface EngineeringSwinburne University of TechnologyHawthornAustralia
  4. 4.Sir Peter MacCallum Department of OncologyThe University of MelbourneParkvilleAustralia
  5. 5.Department of PathologyThe University of MelbourneParkvilleAustralia

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