MECHANISM OF CONCENTRATION QUENCHING OF A XANTHENE DYE ENCAPSULATED IN PHOSPHOLIPID VESICLES

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Abstract
Encapsulating a xanthene dye in phospholipid vesicles produces vesicle solutions that contain dye at very high microscopic concentrations, but have a low overall optical density, thereby eliminating reabsorption. Using this system, we have studied the effects of concentration on the fluorescence lifetime of one such dye, sulforhodamine 101. We have observed that the lifetime decreases as a function of encapsulated dye concentration, which is indicative of collisional quenching. The lifetime decreases from 4.5 nsec for sulforhodamine in dilute aqueous solution to 0.69 ns at an encapsulated concentration of 33 mM. The bimolecular rate constant for this event is 2.6 1010M‐1 s‐1, consistent with a diffusion controlled event. However, the quenching constant calculated from changes in intensity is 2.2 1011M‐1 s‐1. Thus, collisional quenching is not the predominant mechanism of quenching. The absorption spectra of dye in vesicles indicate an important contribution from static complex formation. Förster distance calculations indicate that energy transfer can also occur to a significant extent, with a predicted efficiency of transfer of 34% at a dye concentration of only 1 mM