K-FOCUS: Live-cell high-throughput single-cell analysis of foci colocalization. (A) Illustration of the analysis pipeline for the Cellpose-TrackIt module, which incorporates CellPose segmentation into TrackIt (scale bar = 10 µm). (B) Workflow for manual ROI quality control and foci tracking using the TrackIt GUI. (C) A schematic outlining the colocalization criteria, including user-defined inputs, and key outputs. Additionally, it presents a visual representation of a colocalized and non-colocalized track, along with a kernel density plot of track length per cell for both colocalized and non-colocalized tracks.