Testing Fluorescence Lifetime Standards using Two-Photon Excitation and Time-Domain Instrumentation: Fluorescein, Quinine Sulfate and Green Fluorescent Protein

摘要: It is essential for everyone working with experimental science to be certain that their instruments produce reliable results, and fluorescence lifetime experiments, information about standards crucial. A large part of the literature on dates back 1970s 1980s, use newer faster measuring devices may deem these results unreliable. We have tested three commonly used fluorophores fluorescein, quinine sulfate green fluorescent protein suitability serve as standards, especially two-photon excitation measurements in time-domain. measured absorption emission spectra determine optimal wavelengths detector settings. Fluorescence lifetimes were different concentrations, ranging from 10− 3 − 10− 5 M, well various solvents. Fluorescein was soluble both ethanol, methanol sulfuric acid, while only acid. Green prepared a commercial Tris-HCl, EDTA solution, all produced stable low uncertainties. No siginificant variation concentration any fluorophores, showed single-exponential decays. All carried out using data obtained time-domain time-correlated single-photon counting.