Shannon, M. J.; Eisman, S. E.; Lowe, A. R.; Sloan, T.; Mace, E. M.

Advances in imaging, cell segmentation, and cell tracking now routinely produce microscopy datasets of a size and complexity comparable to transcriptomics or proteomics. New tools are required to process this phenomics type data. Cell PLasticity Analysis TOol (cellPLATO) is a Python-based analysis software designed for measurement and classification of diverse cell behaviours based on clustering of parameters of cell morphology and motility. cellPLATO is used after segmentation and tracking of cells from live cell microscopy data. Morphokinetic information about each cell from each timepoint is used to segregate cells automatically with UMAP/HDBSCAN into behavioural subtypes. Analysis of behavioural sequences and trajectories subsequently generates a phenotypic fingerprint for each condition and representative cells are mathematically identified and graphically displayed for intuitive understanding of each subtype. Here, we use cellPLATO to investigate the role of IL-15 in modulating NK cell migration on ICAM-1 or VCAM-1. We find 8 behavioural subsets of NK cells based on their shape and migration dynamics, and 4 trajectories of behaviour. Therefore, using cellPLATO we show that IL-15 increases plasticity between cell migration behaviours and that different integrin ligands induce different forms of NK cell migration.