Using Biotinylated Fluorophores Attached to Streptavidin Adsorbed to Nanopatterned Gold Surfaces to Enhance Fret

Surface plasmons generated on nanopatterned gold substrates can be used to enhance Förster Resonance Energy Transfer (FRET) between two fluorescent molecules by increasing the local density of optical states. Previously in the Steele lab, DNA with fluorescent molecules along its backbone were attached to the surface of these nanopatterned gold substrates in an effort to precisely space the fluorescent molecules and ascertain the specifications of FRET enhancement. However, this work continually gave results inconsistent with current understanding of fluorescence enhancement. It was hypothesized that these inconsistencies might be due to the fluorescent molecules on the DNA not having proper placement, either due to the DNA not standing upright or otherwise not behaving as we expected. To remedy these issues, this thesis explores using streptavidin and biotin to attach fluorescent molecules to gold nanopatterned surfaces. A procedure was developed to adsorb a single layer of streptavidin to both gold and glass to which a biotinylated target molecule can then be attached. Future work will investigate the feasibility of this method for FRET enhancement.