Robust and automated detection of subcellular morphological motifs in 3D microscopy images

Rapid developments in live-cell 3D microscopy enable imaging of cell morphology and signaling with unprecedented detail. However, tools to systematically measure and visualize the intricate relationships between intracellular signaling, cytoskeletal organization, and downstream cell morphological outputs do not exist. Here we introduce u-shape3D, a computer graphics and machine-learning pipeline to probe molecular mechanisms underlying 3D cell morphogenesis and to test the intriguing possibility that morphogenesis itself affects intracellular signaling. We demonstrate a generic morphological motif detector that automatically finds lamellipodia, filopodia, blebs, and other motifs. Combining motif detection with molecular localization, we measure the differential association of PIP(2) and Kras(V12) with blebs. Both signals associate with bleb edges, as expected for membrane-localized proteins, but only PIP(2) is enhanced on blebs. This indicates that sub-cellular signaling processes are differentially modulated by local morphological motifs. Overall, our computational workflow enables the objective, 3D analysis of the coupling of cell shape and signaling.