title: Divergent lymphocyte signalling revealed by a powerful new tool for analysis of time-lapse microscopy.
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.
authors.affiliation: Centre for Micro-Photonics, Faculty of Engineering and Industrial Sciences, Swinburne University of Technology, Hawthorn, Adelaide, Australia.
authors.forename: Kim
authors.initials: K
authors.lastname: Pham
authors.forename: Raz
authors.initials: R
authors.lastname: Shimoni
authors.forename: Mandy J
authors.initials: MJ
authors.lastname: Ludford-Menting
authors.forename: Cameron J
authors.initials: CJ
authors.lastname: Nowell
authors.forename: Pavel
authors.initials: P
authors.lastname: Lobachevsky
authors.forename: Ze'ev
authors.initials: Z
authors.lastname: Bomzon
authors.forename: Min
authors.initials: M
authors.lastname: Gu
authors.forename: Terence P
authors.initials: TP
authors.lastname: Speed
authors.forename: C Jane
authors.initials: CJ
authors.lastname: McGlade
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Russell
mesh_terms: D000818:Animals; D016764:Cell Polarity / Q000276:immunology; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods*; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology*; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D009419:Nerve Tissue Proteins / Q000276:immunology; D009928:Organ Specificity / Q000276:immunology; D010766:Phosphorylation / Q000276:immunology; D011493:Protein Kinase C / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology*
chemical_list: D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C106293:Numb protein, mouse; C115015:PKC-3 protein; D011493:Protein Kinase C