Electrochemically modulated surface plasmon waves for characterization and interrogation of DNA-based sensors

This work reports on a comparative analysis of electrical and optical measurements for structural characterization and for assessing signal transduction performance of a redox-labeled DNA-based sensing platform. We conducted complementary investigations employing conventional electrochemical techniques with electric current measurements in cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) and confronted those results with optical measurements using surface plasmon waves while the redox assembly was undergoing similar electrochemical modulation as in the electrical CV and EIS measurements. The specific sensor configuration deployed here was composed of a methylene blue (MB)-modified single-stranded DNA (ssDNA) signaling probe and an unlabeled capture ssDNA probe that complements the signaling probe. Two types of signaling probes were employed: one with MB attached to the 3′ end, which positions the redox marker closer to the electrode surface upon hybridization with the capture probe, and the other with MB attached to the 5′ end, which places the redox marker farther from the electrode surface. For each molecular assembly and for each electrochemical modulation protocol, both the electrical and optical experimental data were quantitatively analyzed to determine the surface density of electro-active species and the rate of electron transfer between the redox marker and the electrode surface. Our experimental results highlight the consistency of the confronted methodologies and indicate that optical impedance spectroscopy utilizing electrochemically modulated surface plasmon waves, which is a transduction protocol immune from non-faradaic interferents that invariably are present in the electrical methodology, can provide a powerful route for developing a redox-labeled DNA-hybridization biosensing strategy.