Methods for analysis of time-resolved fluorescence data

We report the results of applying several deconvolution methods to estimate fluorescence decay curves and fluorescence lifetimes. The methods are applied to time-resolved fluorescence signals measured with a homemade spectrofluorometer. We measured the fluorescence emitted by three substances with known lifetimes: fluoresceine (4.03 ns), rhodamine 6G (4.08 ns), and rhodamine B (1.68 ns). Also we measured the fluorescence emitted by three mixtures of two substances. We review three deconvolution methods: Fourier transform, least-squares iterative reconvolution, and Laguerre functions. to estimate the intrinsic fluorescence of the samples. A program was developed in LabView 7.1 to apply the three deconvolution methods from the observed fluorescence signals and the instrumental response signal. We conclude from data analysis results, that deconvolution by Laguerre expansion provides the best approximation of fluorescence lifetimes, producing an error less than 0.8% for substances, and less than 2.5% for mixtures.