A Correction Method for Fluorescence Reduction Using Time-Resolved Fluorometry and Spectrophotometry

Abstract

A method to correct fluorescence reduction due to foreign materials and temperature using time-resolved fluorometry and spectrophotometry has been developed. The factors taken into consideration were dynamic quenching and inner filter effects on the excitation beam and the fluorescence, and the fluorescence intensity with respect to these factors was theoretically analyzed. A theoretical approach for correcting the fluorescence intensity was obtained, with the use of the fluorescence lifetime, the absorbance at the excitation wavelength, and the absorbance at the fluorescence wavelength. Since these parameters are observable, a novel method can be applied to samples in cases where foreign materials and temperature are unknown. The proposed correction method is also suitable for noncontact and remote measurements. From experiments using a UO₂²⁺ ion, the proposed method was verified.