A Divide-and-Conquer Strategy for Quantification of Light Absorption, Scattering, and Emission Properties of Fluorescent Nanomaterials in Solutions.

Optical properties of fluorescent materials including their UV-vis absorption, scattering, and on-resonance fluorescence activities are strongly wavelength-dependent. Reported herein is a divide-and-conquer strategy for experimental quantification of fundamental optical constants of fluorescent nanomaterials including their UV-vis absorption, scattering, and on-resonance-fluorescence (ORF) cross-section spectra and ORF fluorescence and light scattering depolarization spectra. The fluorophore UV-vis extinction spectrum is first divided into a blue and a red wavelength region. The UV-vis extinction cross-section spectrum in the blue wavelength region is decomposed into its absorption and scattering extinction spectra straightforwardly using the established polarized resonance synchronous spectroscopic technique. In its red wavelength region, however, the fluorophores can be simultaneous photon absorbers, scatterers, and anti-Stokes-shifted, on-resonance, and Stokes-shifted fluorescence emitters under the resonance excitation and detection conditions. A polarized anti-Stokes'-shifted, on-resonance, and Stokes'-shifted spectroscopic method is developed for quantifying fluorophore absorption, scattering, one-resonance fluorescence (ORF) cross-section spectra, and scattering and ORF fluorescence depolarization spectra in this wavelength region. Example applications of the presented techniques were demonstrated with fluorescent polystyrene nanoparticles, fluorescent quantum dots, and molecular fluorophores Rhodamine 6G and Eosin Y.