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A microstructured graphitic 4x4 multielectrode array was embedded in a singlecrystal diamond substrate (4x4 {uG-SCD MEA) for real-time monitoring ofexocytotic events from cultured chromaffin cells and adrenal slices. Thecurrent approach relies on the development of a parallel ion beam lithographictechnique, which assures the time effective fabrication of extended arrays withreproducible electrode dimensions. The reported device is suitable forperforming amperometric and voltammetric recordings with high sensitivity andtemporal resolution, by simultaneously acquiring data from 16 rectangularlyshaped microelectrodes (20x3.5 um^2) separated by 200 um gaps. Taking advantageof the array geometry we addressed the following specific issues: i) detectboth the spontaneous and KCl-evoked secretion simultaneously from severalchromaffin cells directly cultured on the device surface, ii) resolve thewaveform of different subsets of exocytotic events, iii) monitoring quantalsecretory events from thin slices of the adrenal gland. The frequency ofspontaneous release was low (0.12 Hz and 0.3 Hz respectively for adrenal slicesand cultured cells) and increased up to 0.9 Hz after stimulation with 30 mM KClin cultured cells. The spike amplitude as well as rise and decay time werecomparable with those measured by carbon fiber microelectrodes and allowed toidentify three different subsets of secretory events associated to "fullfusion" events, "kiss and-run" and "kiss-and-stay" exocytosis, confirming thatthe device has adequate sensitivity and time resolution for real-timerecordings. The device offers the significant advantage of shortening the timeto collect data by allowing simultaneous recordings from cell populationseither in primary cell cultures or in intact tissues.

analytical chemistry

Microelectrode arrays of diamond-insulated graphitic channels for real  time detection of exocytotic events from cultured chromaffin cells and slices  of adrenal glands