Fluorescence anisotropy (FA) of anionic dyes bound to ionic and zwitterionic micelles

Anionic fluorescein and 8-hydroxy-1,3,6-pyrenetrisulfonate (POH), bind to cationic and zwitterionic micelles, experience hindered rotation and exhibit fluorescence anisotropy (FA). Fluorescein emits three lines from its dianion, carboxylate, and phenolate forms. POH emissions are from excited POH* and its deprotonated form, PO^-. Fluorescence was excited by vertically polarized (V) light. Spectra recorded with vertical (IVV) and horizontal (IVH) polarizers in the emitted beam were corrected for instrument response and polarization bias. Corrected line shapes were fit to Gaussians availing the computationally derived second harmonic for better fit precision. FA of the individual forms of the same dye was calculated from the IVV and IVH intensities of each component line. For fluorescein, FA phenolate > carboxylate > dianion and FA PO^-> POH*. Micelle-bound dye conformations, consistent with this order, are presented. Distinguishing between FA of different forms is novel and significant to elucidation of dye-host interactions.