Real-time visualization of intracellular hydrodynamics in single living cells

Abstract

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.

Implications of a poroelastic cytoplasm for the dynamics of animal cell shape

“…Unfortunately, reality is more complex, since both water and ions can cross the plasma membrane rapidly. We know this from responses to changes in external osmolarity [50] (figure 4), and from direct observation of H 2 O for D 2 O exchange, which was followed using CARS microscopy, revealing a timescale of ~2sec [51]. The transporters that promote rapid movement of water and ions across the relatively impermeable lipid bilayer are discussed below.…”

Coherent Anti-Stokes Raman Scattering Correlation Spectroscopy: Probing Dynamical Processes with Chemical Selectivity

“…For instance, CARS microscopy was applied to visualizing intracellular hydrodynamics by use of a line-scanning scheme. 10 High-speed imaging of unstained cells undergoing apoptosis and chromosome distribution in mitotic cells has been carried out using a laser-scanning CARS microscope at an acquisition rate of several seconds per frame. 6 CARS microscopy opens up exciting possibilities of visualizing dynamical changes in living cells and tissues with chemical selectivity, complementary to multiphoton fluorescence microscopy.…”

Comparison of coherent and spontaneous Raman microspectroscopies for noninvasive detection of single bacterial endospores

“…CARS spectroscopy has provided a useful spectroscopic technique for the past 40 years, despite inherent difficulties as discussed below. However, recent years have witnessed renaissance of interest in CARS spectroscopy (5)(6)(7)(8)(9)(10)(11)(12)(13). This recent progress is driven mostly by technical developments in lasers, optics, electronics and computers, which facilitate the widespread use of this promising spectroscopic tool.…”