Evaluation of nanosecond pulse–fluorometry measurements—no need for the excitation function

Abstract

The standard method of evaluating pulse–fluorometry measurements is to deconvolute the data using an experimentally determined excitation function. However, it is known that its determination may be inaccurate, because the instrumental response and the form of the exciting light pulse do vary with the wavelength. This may limit the accuracy of deconvolution and the confidence in its results. Therefore, in this paper an alternative evaluation method is suggested. The excitation function is not needed, if two independent measurements differing in the kinetics of the involved decays, but not in the excitation and emission wavelengths, can be performed. Those two measurements are sufficient to obtain the parameters of both decays.