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Scanning electrochemical microscopy combined with surface plasmon resonance: studies of localized film thickness variations and molecular conformation changes.

The combination of scanning electrochemical microscopy (SECM) with surface plasmon resonance (SPR) is described. By oxidizing ferrocenylalkanethiol self-assembled monolayer (SAM) with SECM-generated Ce4+, the coupled technique, SECM-SPR, is shown to be viable for determining local variations in thin film thickness. Factors (tip/substrate distance, tip potential scan rate, and solution composition change) affecting the SECM-SPR response and operation are also discussed. The approach was further extended to the determination of conformational changes of cytochrome c molecules attached electrostatically onto a negatively charged SAM during its reduction by the tip-generated methyl viologen monocation. The high sensitivity of the SPR equipped with a bicell detector facilitates the measurement of infinitesimal film thickness changes accompanying redox reactions, while the SECM provides a means to obviate the necessity of applying a potential to the SPR substrate, which tends to cause unwanted interferences and complications. The approach also affords an avenue for determining film thickness variations that are not subject to certain effects, such as the surface charge, the heterogeneity of the substrate, and the distance between the redox center of the immobilized molecule and the underlying substrate electrode.