Measurement of very short fluorescence lifetimes by single-photon counting

Abstract

Measurement of very short fluorescence lifetimes by the single‐photon technique is made possible by an improved fluorescence lifetime system. Fluorescence lifetimes of 4.94±0.07 nsec for anthracene in cyclohexane, 640±30 psec for diphenyl butadiene in cyclohexane, and 90±30 psec for erythrosin in water were determined. The use of a small wavelength shift between excitation and emission minimizes the effect of the wavelength dependence of the photomultiplier response and light pulser emission. The effects of deaeration of solutions and time averaging of the excitation profile are presented. We investigated the origin of small‐amplitude early and late artifactual peaks in the light pulser and fluorescence profiles. Complications in the analysis of lifetime data introduced by intrinsic fluorescence and phosphorescence processes in commonly used absorption filters are discussed. Certain ’’blind spots’’ are found in the electronic pulse pileup rejection schemes most commonly used in photon counting.