Metal-Enhanced Fluorescence of Phycobiliproteins from Heterogeneous Plasmonic Nanostructures

Abstract

We report here the use of plasmonic metal nanostructures in the form of silver island films (SiFs) to enhance the fluorescence emission of five different phycobiliproteins. Our findings clearly show that the phycobiliproteins display up to a 9-fold increase in fluorescence emission intensity, with a maximum 7-fold decrease in lifetime when they are assembled as a monolayer above SiFs, as compared to a monolayer assembled on the surface of amine-terminated glass slides of the control sample. The study was also repeated with a thin liquid layer of the phycobiliproteins sandwiched between two glass substrates (and a SiFs and a glass substrate) clamped together. Similarly, the results show a maximum 10-fold increase in fluorescence emission intensity coupled with a 2-fold decrease in lifetime of the phycobiliproteins in the SiF-glass setup as compared to the glass control sample, implying that near-field enhancement of phycobiliprotein emission can be attained both with and without chemical linkage of the proteins to the SiFs. Hence, our results clearly show that metal-enhanced fluorescence (MEF) can potentially be employed to increase the sensitivity and detection limit of the plethora of bioassays that employ phycobiliproteins as fluorescence labels, such as in fluoro-immunoassays where the assay can be tethered on the surface of SiFs, and also in flow cytometry where analytes in the liquid phase could potentially flow through channels coated with SiFs without actually being attached to the silver.