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Magnetron sputtered diamond-like carbon microelectrodes for on-chip measurement of quantal catecholamine release from cells
Authors:Yuanfang Gao  Xiaohui Chen  Sanju Gupta  Kevin D. Gillis  Shubhra Gangopadhyay
Affiliation:Department of Electrical and Computer Engineering, University of Missouri-Columbia, 243 Engineering Bldg. West, Columbia, MO, 65211, USA.
Abstract:Carbon electrodes are widely used in electrochemistry due to their low cost, wide potential window, and low and stable background noise. Carbon-fiber electrodes (CFE) are commonly used to electrochemically measure "quantal" catecholamine release via exocytosis from individual cells, but it is difficult to integrate CFEs into lab-on-a-chip devices. Here we report the development of nitrogen doped diamond-like carbon (DLC:N) microelectrodes on a chip to monitor quantal release of catecholamines from cells. Advantages of DLC:N microelectrodes are that they are batch producible at low cost, and are harder and more durable than graphite films. The DLC:N microelectrodes were prepared by a magnetron sputtering process with nitrogen doping. The 30 mum by 40 mum DLC:N microelectrodes were patterned onto microscope glass slides by photolithography and lift-off technology. The properties of the DLC:N microelectrodes were characterized by AFM, Raman spectroscopy and cyclic voltammetry. Quantal catecholamine release was recorded amperometrically from bovine adrenal chromaffin cells on the DLC:N microelectrodes. Amperometric spikes due to quantal release of catecholamines were similar in amplitude and area as those recorded using CFEs and the background current and noise levels of microchip DLC:N electrodes were also comparable to CFEs. Therefore, DLC:N microelectrodes are suitable for microchip-based high-throughput measurement of quantal exocytosis with applications in basic research, drug discovery and cell-based biosensors.
Keywords:Diamond-like carbon  Magnetron sputtering  Nitrogen doping  Exocytosis  Amperometry  BioMEMS  Lab-on-a-chip
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