Polystyrene Microspheres in Tissue-Simulating Phantoms Can Collisionally Quench Fluorescence

Tissue-simulating phantoms that replicate intrinsic optical properties in a controlled manner are useful for quantitative studies of photon transport in turbid biological media. In such phantoms, polystyrene microspheres are often used to simulate tissue optical scattering. Here, we report that using polystyrene microspheres in fluorescent tissue-simulating phantoms can reduce fluorophore quantum yield via collisional quenching. Fluorescence lifetime spectroscopy was employed to characterize quenching in phantoms consisting of a fluorescein dye and polystyrene microspheres (scattering coefficients _s 100-600cm^–1). For this range of tissue-simulating phantoms, analysis using the Stern-Volmer equation revealed that collisional quenching by polystyrene microspheres accounted for a decrease in fluorescence intensity of 6-17% relative to the intrinsic intensity value when no microspheres (quenchers) were present. The intensity decrease from quenching is independent of additional, anticipated losses arising from optical scattering associated with the microspheres. These results suggest that quantitative fluorescence measurements in studies employing such phantoms may be influenced by collisional quenching.