We have developed a wide-field time-resolved imaging system to image quantitatively both the fluorescence lifetime and the rotational correlation time of a fluorophore. Using a polarization-resolved imager, we simultaneously image orthogonal polarization components of the fluorescence emission onto a time-gated intensified CCD. We demonstrate imaging of solvent viscosity variations through the rotational correlation time of fluorescein in a multiwell plate and apply this technique to probe the microviscosity in live cells.
© 2004 Optical Society of America
(000.1430) General : Biology and medicine
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(300.2530) Spectroscopy : Fluorescence, laser-induced
(300.6500) Spectroscopy : Spectroscopy, time-resolved
Klaus Suhling*, Jan Siegel, Peter M. P. Lanigan, Sandrine Lévêque-Fort, Stephen E. D. Webb**, David Phillips, Daniel M. Davis, and Paul M. W. French, "Time-resolved fluorescence anisotropy imaging applied to live cells," Opt. Lett. 29, 584-586 (2004)