Steady-state and time-resolved fluorescence spectroscopy of F2N12S in DOPC/Chol LUVs. (A) Steady-state spectra of F2N12S in DOPC/Chol LUVs with increasing fractions of Chol: 0.07 (black), 0.14 (red), 0.25 (magenta), and 0.32 (orange). Spectra of F2N12S in DOPC and SM/Chol are in green and blue, respectively. The spectra are normalized at the T∗ band maximum. (Inset) Dependence of the hydration parameter of F2N12S on the Chol fraction in DOPC/Chol LUVs. The probe hydration in DOPC was taken as 100%. (B) Time-resolved fluorescence parameters of F2N12S in DOPC/Chol LUVs with increasing Chol fraction. Experiments were performed in cuvettes. Color code as in (A). The ∼30 ps component describing the ESIPT reaction was omitted in the graph. Emission was recorded at 590 nm. To see this figure in color, go online.