A method for fabricating submicrometer-sized gold electrodes of conical or spherical geometry is described. By generating an electric arc between an etched gold microwire and a tungsten counter electrode, the very end of the gold microwire can be melted and given an overall spherical or conical shape a few hundred nanometers in size. The whole wire is subsequently insulated via the cathodic deposition of electrophoretic paint. By applying a high-voltage pulse to the microwire, the film covering its very end can then be selectively removed, thus exposing a submicrometer-sized electrode surface of predefined geometry. The selective exposure of the preformed end of the microwire is demonstrated by cyclic voltammetry, scanning electron microscopy, and metal electrodeposition experiments. The electrophoretic paint coating provides a low-capacitance, robust insulating film allowing exploration of a very wide potential window in aqueous solution. The submicrometer-sized electrodes can easily be turned into probes suitable for combined scanning electrochemical-atomic force microscopy by bending and flattening the gold microwire so that the tip is borne by a flexible enough arm. The good agreement between theoretical and experimental scanning electrochemical microscopy approach curves thus obtained confirms that only the very end of the tip, of predefined geometry, is exposed to the solution.

The proof-of-principle of a nonoptical real-time PCR method based on the electrochemical monitoring of a DNA intercalating redox probe that becomes considerably less easily electrochemically detectable once intercalated to the amplified double-stranded DNA is demonstrated. This has been made possible thanks to the finding of a redox intercalator that (i) strongly and specifically binds to the amplified double-stranded DNA, (ii) does not significantly inhibit PCR, (iii) is chemically stable under PCR cycling, and (iv) is sensitively detected by square wave voltammetry during PCR cycling. Among the different DNA intercalating redox probes that we have investigated, namely, methylene blue, Os[(bpy)(2)phen](2+), Os[(bpy)(2)DPPZ](2+), Os[(4,4'-dimethyl-bpy)(2)DPPZ](2+) and Os[(4,4'-diamino-bpy)(2)DPPZ](2+) (with bpy = 2,2'-bipyridine, phen = phenanthroline, and DPPZ = dipyrido[3,2-a:2',3'-c]phenazine), the one and only compound with which it has been possible to demonstrate the proof-of-concept is the Os[(bpy)(2)DPPZ](2+). In terms of analytical performances, the methodology described here compares well with optical-based real-time PCRs, offering finally the same advantages than the popular and routinely used SYBR Green-based real-time fluorescent PCR, but with the additional incomes of being potentially much cheaper and easier to integrate in a hand-held miniaturized device.