Time-resolved fluorescence anisotropy imaging applied to live cells.
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.
Suhling
Klaus
K
Departments of Physics, Biological Sciences and Chemistry, Imperial College London, London SW7 2BW, UK. k.suhling@imperial.ac.uk
Siegel
Jan
J
Lanigan
Peter M P
PM
Lévêque-Fort
Sandrine
S
Webb
Stephen E D
SE
Phillips
David
D
Davis
Daniel M
DM
French
Paul M W
PM
D016475:3T3 Cells; D000818:Animals; D016880:Anisotropy; D002468:Cell Physiological Phenomena; D004285:Dogs; D004867:Equipment Design; D005453:Fluorescence; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D008859:Microscopy, Polarization / Q000295:instrumentation; D008962:Models, Theoretical; D012617:Sea Urchins; D013997:Time Factors; D014783:Viscosity