Divergent lymphocyte signalling revealed by a powerful new tool for analysis of time-lapse microscopy

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Title: Divergent lymphocyte signalling revealed by a powerful new tool for analysis of time-lapse microscopy
Author(s): Pham, Kim; Shimoni, Raz; Ludford-Menting, Mandy J.; Nowell, Cameron J.; Lobachevsky, Pavel; Bomzon, Ze'ev; Gu, Min; Speed, Terence P.; McGlade, C. Jane; Russell, Sarah M.
Abstract: We describe a new approach for interactive analysis of time-lapse microscopy, and apply this approach to elucidating whether polarity regulation is conserved between epithelial cells and lymphocytes. A key advantage of our analysis platform, 'TACTICS', is the capacity to visualize individual data points in the context of large data sets, similar to standard approaches in flow cytometry. Scatter plots representing microscopic parameters or their derivations such as polarity ratios are linked to the original data such that clicking on each dot enables a link to images and movies of the corresponding cell. Similar to flow cytometric analysis, subsets of the data can be gated and reanalyzed to explore the relationships between different parameters. TACTICS was used to dissect the regulation of polarization of the cell fate determinant, Numb, in migrating lymphocytes. We show here that residues of Numb that are phosphorylated by atypical protein kinase C (aPKC) to mediate apicobasal polarity in epithelial cells are not required for polarization of Numb in T cells, indicating that the role of aPKC is not conserved between lymphocytes and epithelia.
Publication type: Journal article
Research centre: Swinburne University of Technology. Faculty of Engineering and Industrial Sciences. Centre for Micro-Photonics
Source: Immunology and Cell Biology, Vol. 91, no. 1 (Jan 2013), pp. 70-81
Publication year: 2013
FOR Code(s): 0601 Biochemistry and Cell Biology; 1107 Immunology
Keyword(s): Cell polarity; Epithelial cells; Image analysis; Lymphocytes; Numb; Polarity regulation; Protein kinase C; TACTICS; T-cell migration; Time-lapse microscopy
Publisher: Nature Publishing Group
ISSN: 0818-9641
Publisher URL: http://dx.doi.org/10.1038/icb.2012.49
Additional information: The authors acknowledge support from the Australian National Health and Medical Research Council (NHMRC), the Human Frontiers Science Programme, the Australian Research Council (ARC) and the Australian Cancer Research Foundation (ACRF).
Peer reviewed