The detection of quantal exocytic events from neurons and neuroendocrine cells is a challenging task in neuroscience. One of the most promising platforms for the development of a new generation of biosensors is diamond, due to its biocompatibility, transparency and chemical inertness. Moreover, the electrical properties of diamond can be turned from a perfect insulator into a conductive material (resistivity ~mΩ·cm) by exploiting the metastable nature of this allotropic form of carbon. A 16‑channels MEA (Multi Electrode Array) suitable for cell culture growing has been fabricated by means of ion implantation. A focused 1.2 MeV He+ beam was scanned on a IIa single-crystal diamond sample (4.5 × 4.5 × 0.5 mm3) to cause highly damaged sub-superficial structures that were defined with micrometric spatial resolution. After implantation, the sample was annealed. This process provides the conversion of the sub-superficial highly damaged regions to a graphitic phase embedded in a highly insulating diamond matrix. Thanks to a three-dimensional masking technique, the endpoints of the sub-superficial channels emerge in contact with the sample surface, therefore being available as sensing electrodes. Cyclic voltammetry and amperometry measurements of solutions with increasing concentrations of adrenaline were performed to characterize the biosensor sensitivity. The reported results demonstrate that this new type of biosensor is suitable for in vitro detection of catecholamine release.
Diamond applications potential for biosensing devices have been highlighted by several authors, especially concerning the long-term stability of covalent functionalisations on its surface. Additionally, in electrochemistry boron doped diamond electrodes (NA ∼ 1020 cm3) show high corrosion resistance and a large hydrolysis window. These features, recognised and exploited in industrial applications, have up to now found little resonance in the life-sciences. Here we present diamond microelectrode arrays based on (1) nanocrystalline diamond (NCD) thin films and (2) single crystal diamond (SCD). NCD is necessary for large area applications like arrays, but graphitic grain boundaries may influence its behaviour. The ideal case SCD is covered here for comparison. The array design consists of four electrodes whose sensitive area is delimited by means of a patterned photoresist. Two different patterns were used to realise a layout with four independent openings (15 µm diameter) for simultaneous detection on multiple cells and a layout with one single window (25 µm diameter) intersecting all four electrodes to create a quadrupolar detector suitable for mapping the activity of single cells. Early results validated the suitability of both NCD and SCD devices: (1) cyclic-voltammetry measurements confirmed the adrenaline oxidation potential on the presented microelectrodes around 650 mV; (2) alternating applications of 1 mM adrenaline and saline rinsing solutions showed negligible electrode fouling; and (3) interfaced to single adrenal chromaffin cells, the devices clearly detected sustained sequences of quantal events (10–100 pA amplitude, 50–100 ms duration) associated to the vesicular release of adrenaline and noradrenaline during exocytosis induced by cell-depolarisation.