Rapid Single-Cell Measurement of Transient Transmembrane Water Flow under Osmotic Gradient

While aquaporin (AQP) gating dynamically regulates transmembrane water permeability for cellular homeostasis, its mechanisms remain poorly understood compared to ion channels. A central challenge is the lack of methods to measure water flow through AQPs with the spatiotemporal resolution and sensitivity equivalent to patch-clamp recordings of ion fluxes, a limitation stemming from the electrically silent nature of water transport. We introduce a technique to rapidly detect cytoplasmic flows induced by osmotic-gradient-driven transmembrane water transport in single adherent human cancer cells. This approach enables direct measurement of AQP-mediated water transport and provides a powerful tool to investigate AQP function and regulation and cytoplasmic flow dynamics at the single-cell level.