A Molecular Fluorescence Spectrometer That Corrects for the Effects of Quenching

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Abstract
A fluorescence spectrometer has been constructed that simultaneously determines fluorescence intensity and excited-state lifetime. The ratio of intensity to lifetime is used to correct for the reduction in fluorescence intensity caused by collisional quenching. A frequency-domain technique is used to determine the excited-state lifetime in real-time. Computer-driven data acquisition provides instantaneous readout of the fluorescence intensity, the excited-state lifetime, and the ratio of these quantities. Correction for quenching by oxygen is demonstrated.