Real-time visualization of intracellular hydrodynamics in single living cells

• DOI: 10.1073/pnas.98.4.1577
• Publication Date: 2012-07-26T23:12:10Z
• Authors: Eric O. Potma; Wim P. de Boeij; Peter J. M. van H...; Douwe A. Wiersma
• Keywords: Intracellular Fluid; 0303 health sciences; Cell Membrane Permeability; Time Factors; Water; Naphthalenes; NMR; Diffusion; 03 medical and health sciences; COS Cells; Chlorocebus aethiops; Image Processing, Computer-Assisted; Animals; Dictyostelium; DIFFUSIONAL WATER PERMEABILITY; Deuterium Oxide; Fluorescent Dyes

Intracellular water concentrations in single living cells were visualized by nonlinear coherent anti-Stokes Raman scattering (CARS) microscopy. In combination with isotopic exchange measurements, CARS microscopy allowed the real-time observation of transient intracellular hydrodynamics at a high spatial resolution. Studies of the hydrodynamics in the microorganism Dictyostelium discoideum indicated the presence of a microscopic region near the plasma membrane where the mobility of water molecules is severely restricted. Modeling the transient hydrodynamics eventuated in the determination of cell-specific cytosolic diffusion and plasma membrane permeability constants. Our experiments demonstrate that CARS microscopy offers an invaluable tool for probing single-cell water dynamics.