Direct visualization and quantitative analysis of water diffusion in complex biological tissues using CARS microscopy.
To date, it has not been possible to measure microscopic diffusive water movements in epithelia and in the interstitial space of complex tissues and organs. Diffusive water movements are essential for life because they convey physiologically important small molecules, e.g. nutrients and signaling ligands throughout the extracellular space of complex tissues. Here we report the development of a novel method for the direct observation and quantitative analysis of water diffusion dynamics in a biologically organized tissue using Coherent Anti-Stokes Raman Scattering (CARS) microscopy. Using a computer simulation model to analyze the CARS O-H bond vibration data during H2O/D2O exchange in a 3D epithelial cyst, we succeeded in measuring the diffusive water permeability of the individual luminal and basolateral water pathways and also their response to hormonal stimulation. Our technique will be applicable to the measurement of diffusive water movements in other structurally complex and medically important tissues and organs.
1] Department of Pharmacology, Keio University School of Medicine, Tokyo, Japan [2].
Ying-Chun
YC
Yu
Yoshiro
Y
Sohma
Shinichi
S
Takimoto
Takayuki
T
Miyauchi
Masato
M
Yasui
D000818:Animals; D051401:Aquaporin 4 / Q000378:metabolism; D002463:Cell Membrane Permeability; D041961:Deuterium Exchange Measurement; D004058:Diffusion; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008853:Microscopy / Q000379:methods*; D008954:Models, Biological; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted; D013059:Spectrum Analysis, Raman; D014162:Transfection; D014667:Vasopressins / Q000494:pharmacology; D014867:Water / Q000378:metabolism*
D051401:Aquaporin 4; D014867:Water; D014667:Vasopressins