Cell geometry distinguishes migration‐associated heterogeneity in two‐dimensional systems

In vitro migration assays are a cornerstone of cell biology and have found extensive utility in research. Over the past decade, several variations two-dimensional (2D) assay improved our understanding this fundamental process. However, ability these approaches to capture functional heterogeneity during their accessibility inexperienced users has been limited. We downloaded published time-lapse 2D data sets subjected them feature extraction with Fiji software. used “Analyze Particles” tool extract 10 geometry features (CGFs), which were grouped into “shape,” “size,” “position” descriptors. Next, we defined migratory status cells using “MTrack2” plugin. All obtained from further rigorous statistical analysis R version 4.0.2. observed consistent associative trends between size shape descriptors validated observations across four independent sets. identify characterize “nonmigrator (NM)” “migrator (M)” subsets. Statistical allowed us considerable NM subset. Interestingly, differences 2D-packing appeared affect CGF within developed an analytical pipeline open source tools, morphologically subsets label-free, imaging data. Our quantitative approach identified nonmigratory predicted influence on migration.