Real-time Four-dimensional Imaging and Flow Dynamics Analysis of Cilia-driven Transport in Mammalian Tissues

Quantifying cilia-driven transport in mammalian tissues requires resolving rapid ciliary dynamics while preserving native three-dimensional geometry. Here we combine real-time four-dimensional light-field microscopy with tomographic particle tracking and physics-informed volumetric flow reconstruction to simultaneously image ciliary activity and reconstruct three-dimensional velocity fields in minimally dissected tissues. Applied across brain, respiratory, and reproductive epithelia, the workflow reveals recurrent vortical flow structures and enables quantitative analysis of flow-tissue coupling across individuals.