Laser-induced fluorescence (LIF) spectroscopy using dyes is frequently applied for characterization of liquids and two-phase flows. The technique is utilized e.g., for mixing studies, thermometry, or droplet sizing. One major application of the LIF technique combined with Mie-scattering is the planar measurement of droplet sizes in spray systems. However, its uncertainty is determined, among others, by varying dye concentration and temperature changes occurring during mixing and droplet evaporation. Systematic experimental investigations are necessary to determine the influence of dye enrichment effects on the LIF-signal of single droplets. For these investigations, the fluorescence dye Eosin-Y is dissolved in water and ethanol, which are typical solvents and working fluids in bio-medical applications and power engineering. A photo-physical characterization of the mixtures under various conditions was conducted using a spectrometric LIF setup and a micro cell. For ethanol, a small temperature dependency of the Eosin-Y LIF signal is observed up to 373 K. Photo-dissociation of Eosin-Y is negligible for solution in ethanol while it is distinct in water. The LIF signals of the single droplets are studied with an acoustic levitator. Effects of droplet evaporation, droplet deformation and varying dye concentration on the LIF-signal are studied. The single droplet measurements revealed a complex change of the fluorescence signal with reduced droplet size. This is due to droplet deformations leading to variations in the internal illumination field as well as dye enrichment during evaporation.