Abstract

The fluorescence decay analysis of intramolecular two-state excited-state processes with added quencher is discussed in terms of compartments. The kinetic equations specifying the fluorescence decay and the time-course of the two excited-state species concentrations are expressed in terms of the rate constants and the spectroscopic parameters b1 and c1. b1 and c1 are respectively the relative absorbance and the normalized spectral emission weighting factor of species 1. The report investigates what has to be known beforehand to determine all relevant parameters. The results of this identifiability study indicate that the following conditions have to be satisfied in order to make an intramolecular two-state excited-state system with added quencher identifiable. First, at least three different quencher concentrations must be used. Second, the two rate constants of quenching must be different. Third, at least one parameter must be known. This parameter can be (1) one rate constant which is not a rate constant of quenching, (2) one b1 value or, (3) one c1 value. In each of these cases an alternative set of system parameters is mathematically possible. A unique solution is guaranteed when the fluorescence decays of a quenched model compound are included in the compartmental analysis.© (1992) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

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