Real-time visualization of intracellular hydrodynamics in single living cells.
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.
Potma
E
E
Ultrafast Laser and Spectroscopy Laboratory, Materials Science Centre, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
de Boeij
W P
WP
van Haastert
P J
PJ
Wiersma
D A
DA
D000818:Animals; D019556:COS Cells; D002463:Cell Membrane Permeability; D002522:Chlorocebus aethiops; D017666:Deuterium Oxide / Q000737:chemistry*; D004023:Dictyostelium / Q000737:chemistry; D004058:Diffusion; D005456:Fluorescent Dyes; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007424:Intracellular Fluid / Q000737:chemistry; D009281:Naphthalenes; D013997:Time Factors; D014867:Water / Q000737:chemistry*
D005456:Fluorescent Dyes; D009281:Naphthalenes; D014867:Water; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid; D017666:Deuterium Oxide