Journal article
Divergent lymphocyte signalling revealed by a powerful new tool for analysis of time-lapse microscopy
K Pham, R Shimoni, MJ Ludford-Menting, CJ Nowell, P Lobachevsky, Z Bomzon, M Gu, TP Speed, CJ McGlade, SM Russell
Immunology and Cell Biology | NATURE PUBLISHING GROUP | Published : 2013
DOI: 10.1038/icb.2012.49
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 e..
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Funding Acknowledgements
OP9-Dll1 cells were from Juan-Carlos Zuniga-Pflucker (University of Toronto, Canada), and the mCherry-tubulin expression construct was provided by Stephen Ting (Peter MacCallum Cancer Centre). Microfabricated cell paddocks were supplied by Daniel Day (Swinburne University and Microsurfaces Pty Ltd). We thank the MATLAB community for sharing many useful open codes. The work was funded by 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), and supported by the animal, flow cytometry and microscopy core facilities of the PeterMac. We thank members of the Russell Lab, David Izon, Patrick Humbert, Ellen Robey, Sarah Ellis, Joe Trapani and Moshe Olshansky for helpful advice and discussions.