An implantable micromachined multi-electrode array (MEA) microprobe modified for utilization as a complete electrochemical biosensor for rapid glutamate detection is described. A post-fabrication method for electrochemical deposition of an iridium oxide (IrOx) film onto a designated microelectrode enabled incorporation of an IrOx reference electrode (RE) on the microprobe. The on-probe IrOx RE provides an alternative to the commonly utilized Ag/AgCl wire RE, which has been shown to be unstable and to cause an inflammatory response in living tissue. The IrOx film electrodeposited onto a platinum site was tested as part of a complete chemical sensing system that included a platinum counter electrode and enzymatic glutamate sensing electrodes all on a single silicon-based MEA platform. The thin film IrOx was mechanically robust enough to endure conditions of repeated heating and wetting during the MEA fabrication process. The pH dependence of the IrOx open circuit potential (OCP) was measured at -77±0.4 mV/pH and remained stable over a two-week period. The on-probe IrOx RE was tested in a two- and three-electrode system with glutamate biosensors. The biosensors were shown to detect a physiologically relevant range of glutamate concentrations and to reject the interferents, dopamine and ascorbic acid. By incorporating all of the electrodes onto a single device, baseline noise was reduced by an average of ∼61% in vitro and ∼71% in vivo with reduced tissue damage, since only a single probe needed to be implanted.
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