Fluorescence anisotropy controlled by light quenching.
We demonstrated that fluorescence anisotropy can be effectively decreased or increased in the presence of light quenching, depending on relative polarizations of excitation and quenching pulses. For parallel light quenching, anisotropy decreases to 0.103 and z-axis symmetry is preserved. In the presence of perpendicular light quenching, the steady-state anisotropy of a pyridine-2-glycerol solution increases from 0.368 for an unquenched sample to 0.484 for a quenched one. We show that the angular distribution of transition moments loses z-axis symmetry in the presence of perpendicular light quenching. In these cases we used more general definitions of anisotropy. Induced by light quenching, anisotropy can be applied in both steady-state and time-resolved measurements. In particular, the systems with low or no anisotropy can be investigated with the proposed technique.
Gryczynski
I
I
Department of Biochemistry and Molecular Biology, University of Maryland at Baltimore 21201, USA.
Gryczynski
Z
Z
Lakowicz
J R
JR
D005454:Fluorescence Polarization / Q000379:methods*; D007700:Kinetics; D008027:Light; D011725:Pyridines
D011725:Pyridines; C023666:pyridine