Tracking Coordinated Cellular Dynamics in Time-Lapse Microscopy with ARCOS.px

Abstract

Subcellular structures like actin waves, podosomes and focal adhesions exhibit dynamic organization that traditional tracking methods struggle to capture. We developed ARCOS.px, an open-source Python package with an interactive napari plugin, to identify and track spatial clusters in microscopy time-lapse images. Unlike conventional approaches that focus on individual particles or bulk intensity changes, ARCOS.px tracks spatial relationships, coordination, and lineage evolution over time. We demonstrate the method by tracking podosome-like structures and RhoA activity waves triggered by growth factors or optogenetics. We revealed how various drugs affect the stability, mobility, and interactions of podosome-like structures in REF52 cells. Additionally, we discovered that actin waves in these cells are followed by active RhoA and Myosin, suggesting feedback mechanisms that recruit active RhoA at their trailing edge. ARCOS.px can be applied to diverse research contexts requiring the tracking of discontinuous yet dynamically associated regions.