title: Time-resolved fluorescence anisotropy imaging applied to live cells.
abstract: 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.
authors.affiliation: Departments of Physics, Biological Sciences and Chemistry, Imperial College London, London SW7 2BW, UK. k.suhling@imperial.ac.uk
authors.forename: Klaus
authors.initials: K
authors.lastname: Suhling
authors.forename: Jan
authors.initials: J
authors.lastname: Siegel
authors.forename: Peter M P
authors.initials: PM
authors.lastname: Lanigan
authors.forename: Sandrine
authors.initials: S
authors.lastname: Lévêque-Fort
authors.forename: Stephen E D
authors.initials: SE
authors.lastname: Webb
authors.forename: David
authors.initials: D
authors.lastname: Phillips
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Davis
authors.forename: Paul M W
authors.initials: PM
authors.lastname: French
mesh_terms: 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