Comparison of Methods and Achievable Uncertainties for the Relative and Absolute Measurement of Photoluminescence Quantum Yields

The photoluminescence quantum yield (Phi(f)) that presents a direct measure for the efficiency of the conversion of absorbed photons into emitted photons is one of the spectroscopic key parameters of functional fluorophores. It determines the suitability of such materials for applications in, for example, (bio) analysis, biosensing, and fluorescence imaging as well as as active components in optical devices. The reborn interest in accurate Phi(f) measurements in conjunction with the controversial reliability of reported Phi(f) values of many common organic dyes encouraged us to compare two relative and one absolute fluorometric method for the determination of the fluorescence quantum yields of quinine sulfate dihydrate, coumarin 153, fluorescein, rhodamine 6G, and rhodamine 101. The relative methods include the use of a chain of Phi(f) transfer standards consisting of several standard dye versus reference dye pairs linked to a golden Phi(f) standard that covers the ultraviolet and visible spectral region, and the use of different excitation wavelengths for standard and sample, respectively. Based upon these measurements and the calibration of the instruments employed, complete uncertainty budgets for the resulting Phi(f)-values are derived for each method, thereby providing evaluated standard operation procedures for Phi(f) measurements and, simultaneously, a set of assessed Phi(f) standards.