We demonstrate the enhancement of fluorescence emission from a dye, 5-carboxyfluorescein (FAM), which couples with surface plasmons at the spectral channels of excitation and emission. Experiments and calculations revealed that bimetallic (gold-silver) plasmon, as compared to the monometallic ones, allowed such coupling to be enhanced, at both the spectral channels. We achieved the maximum fluorescence enhancement level of 46.5-fold, with markedly high reproducibility (coefficient of variation ~ 0.5%) at a FAM concentration of 10 nM. We also found that higher fluorescence enhancement was more likely to be reproducible. This encourages the use of this technology for practical applications in fluorescence-based biochemical assays. Moreover, we investigated a FAM concentration-dependent enhancement of fluorescence. It was found that fluorescence enhancement decreased and saturated at above 10 nM concentration possibly due to partial photo-bleaching of FAM molecules.
Keywords: 5-carboxyfluorescein; fluorescence enhancement; nano-biophotonics; optical biosensors; surface plasmon resonance.