Synergistic effects of micro/nano modifications on electrodes for microfluidic electrochemical ELISA

Microfluidic electrochemical sensing has been considered to be highly efficient. However, we showed, by using numerical simulations in this study, that a planar electrode formed on the bottom of a microchannel is exposed to only a small fraction of analytes in amperometric detection. We also showed that three-dimensional (3D) micropillar electrodes significantly improve the detection current. The practical performance was evaluated using 3D micropillar electrodes fabricated by photolithography. The output current increased as the diameters of the micropillars decreased, as predicted by the simulations. It is noteworthy that the current enhancements obtained with the 3D electrodes were larger than those expected from an increase in the surface area. Further increase in current was achieved by electrical deposition of nanoporous gold-black onto the surface of the 3D electrode: when a 3D electrode with micropillars 30 μm in diameter was used, the output current was approximately 20 times that obtained with a 2D electrode without modification. The applicability of the micropillar electrodes was demonstrated in electrochemical enzyme-linked immunosorbent assay (ELISA) of bone metabolic marker proteins. Although an increase in the surface area of the electrode leads to more noise in general, there is no significant difference in the signal-to-noise ratio between the modified 3D electrode and the 2D electrode without modification in the ELISA experiments. This nanoporous micropillar electrode could potentially be a useful component for the development of on-site diagnosis systems.