Kinetic spectroscopy of erythrosin phosphorescence and delayed fluorescence in aqueous solution at room temperature.
The photophysics and polarization of the phosphorescence and delayed fluorescence of erythrosin in conditions compatible with the current biological applications of the dye (aqueous buffers at pH 7.4 at ambient temperatures) and in ethanol have been studied as a function of dye concentration (10(-7)-10(-5) M) and temperature (245-333 K). The emission decay is strictly single exponential and the detailed kinetic analysis of all the rate processes connected with the emitting T1 state showed that (1) the lowering of the emission lifetime at the higher temperatures is due to a very efficient self-quenching process, (2) the back intersystem crossing rate T1-->S1 is temperature dependent (delta ETS approximately 7 kcal mol-1) but the T1-->S0 is not (Ea < 0.1 kcal mol-1) and (3) both intersystem crossing processes are very sensitive to solvent polarity, which accounts for the solvent dependence of the phosphorescence yield and lifetime. The high value of the phosphorescence anisotropy (r0 = 0.25 +/- 0.006) is independent of the excitation and emission wavelengths, and its evolution in time accurately reflects the rotational restrictions in solid solutions. The relevance of these findings to studies with protein-dye conjugates is also outlined to facilitate the design and interpretation of phosphorescence depolarization experiments that probe the microsecond-ms dynamics of biomolecules and supramolecular systems.
Duchowicz
R
R
Centro de Investigaciones Opticas (CONICET-CIC BA), University La Plata, Argentina.
Ferrer
M L
ML
Acuña
A U
AU
D004923:Erythrosine / Q000737:chemistry*; D007700:Kinetics; D049449:Luminescence; D010777:Photochemistry; D013050:Spectrometry, Fluorescence / Q000379:methods; D013053:Spectrophotometry / Q000379:methods
D004923:Erythrosine