A rapid excitation-emission matrix fluorometer utilizing supercontinuum white light and acousto-optic tunable filters.

Abstract

Scanning speed and coupling efficiency of excitation light to optic fibres are two major technical challenges that limit the potential of fluorescence excitation-emission matrix (EEM) spectrometer for on-line applications and in vivo studies. In this paper, a novel EEM system, utilizing a supercontinuum white light source and acousto-optic tunable filters (AOTFs), was introduced and evaluated. The supercontinuum white light, generated by pumping a nonlinear photonic crystal fiber with an 800 nm femtosecond laser, was efficiently coupled into a bifurcated optic fiber bundle. High speed EEM spectral scanning was achieved using AOTFs both for selecting excitation wavelength and scanning emission spectra. Using calibration lamps (neon and mercury argon), wavelength deviations were determined to vary from 0.18 nm to -0.70 nm within the spectral range of 500-850 nm. Spectral bandwidth for filtered excitation light broadened by twofold compared to that measured with monochromatic light between 650 nm and 750 nm. The EEM spectra for methanol solutions of laser dyes were successfully acquired with this rapid fluorometer using an integration time of 5 s.