title: Imaging Asymmetric T Cell Division.
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.
authors.affiliation: Faculty of Science, Engineering and Technology, Centre for Micro-Photonics, Swinburne University of Technology, Mail No H74, PO Box 218, Hawthorn, VIC, 3122, Australia.|Immune Signalling Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC, 3002, Australia.|Faculty of Science, Engineering and Technology, Biointerface Engineering, Swinburne University of Technology, Hawthorn, VIC, 3122, Australia.
authors.forename: Mirren
authors.initials: M
authors.lastname: Charnley
authors.affiliation: Faculty of Science, Engineering and Technology, Centre for Micro-Photonics, Swinburne University of Technology, Mail No H74, PO Box 218, Hawthorn, VIC, 3122, Australia. sarah.russell@petermac.org.|Immune Signalling Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC, 3002, Australia. sarah.russell@petermac.org.|Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. sarah.russell@petermac.org.|Department of Pathology, The University of Melbourne, Parkville, VIC, 3052, Australia. sarah.russell@petermac.org.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Russell
mesh_terms: D000818:Animals; D060049:Asymmetric Cell Division / Q000276:immunology*; D017173:Caenorhabditis elegans / Q000276:immunology*; D048749:Cytokinesis / Q000276:immunology*; D004331:Drosophila melanogaster; D008856:Microscopy, Fluorescence / Q000379:methods; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
keywords: Asymmetric cell division; Cell fate determinants; Cell polarity; Fluorescence microscopy; Image quantification; Lymphocytes; T Cells; Thymocyte