Design and Calibration of a Rather Simple Computer-Controlled Pulsed Laser Spectrofluorimeter

Abstract

Recently a series of papers dealt with up-to-date fluorimeters, in particular those utilizing pulsed laser light sources. Design concepts, as well as several advantages for the study of minute amounts of substances, were discussed. Since in many spectrometers pulsed lasers are used as exciting light sources, one should exercise care when choosing the sampling duration of the observed fluorescence; otherwise, the pulsed spectrum can grossly differ from the conventional stationary spectrum. Few authors have made proper spectral corrections in their research findings. In this paper we will describe a simple, straightforward calibration procedure and demonstrate that pulsed spectra are indistinguishable from stationary spectra, provided the sampling time period is comparable with the fluorescence decay time.