Accurate and stable chronic in vivo voltammetry enabled by a replaceable subcutaneous reference electrode

In vivo sensing of neurotransmitters has provided valuable insight into both healthy and diseased brain. However, chronically implanted Ag/AgCl reference electrodes suffer from degradationgradation, resulting in errors in the potential at the working electrode. Here, we report a simple, effective way to protect in vivo sensing measurements from reference polarization with a replaceable subcutaneously implanted reference. We compared a brain-implanted reference and a subcutaneous reference and observed no difference in impedance or dopamine redox peak separation in an acute preparation. Chronically, peak background potential and dopamine oxidation potential shifts were eliminated for three weeks. Scanning electron microscopy shows changes in surface morphology and composition of chronically implanted Ag/AgCl electrodes, and postmortem histology reveals extensive cell death and gliosis in the surrounding tissue. As accurate reference potentials are critical to in vivo electrochemistry applications, this simple technique can improve a wide and diverse assortment of in vivo preparations.