Nanocrystalline diamond microelectrode arrays fabricated on sapphire technology for high-time resolution of quantal catecholamine secretion from chromaffin cells.

The quantal release of oxidizable molecules can be successfully monitored by means of polarized carbon fiber microelectrodes (CFEs) positioned in close proximity to the cell membrane. To partially overcome certain CFE limitations, mainly related to their low spatial resolution and lack of optical transparency, we developed a planar boron-doped nanocrystalline diamond (NCD) prototype, grown on a transparent sapphire wafer. Responsiveness to applied catecholamines as well as the electrochemical and optical properties of the NCD-based device were first characterized by cyclic voltammetry and optical transmittance measurements. By stimulating chromaffin cells positioned on the device with external KCl, well-resolved quantal exocytotic events could be detected either from one NCD microelectrode, or simultaneously from an array of four microelectrodes, indicating that the chip is able to monitor secretory events (amperometric spikes) from a number of isolated chromaffin cells. Spikes detected by the planar NCD device had comparable amplitudes, kinetics and vesicle diameter distributions as those measured by conventional CFEs from the same chromaffin cell.

[1]  J. Galligan,et al.  In vitro continuous amperometry with a diamond microelectrode coupled with video microscopy for simultaneously monitoring endogenous norepinephrine and its effect on the contractile response of a rat mesenteric artery. , 2006, Analytical chemistry.

[2]  Michele Dipalo,et al.  Combining diamond electrodes with GaN heterostructures for harsh environment ISFETs , 2009 .

[3]  C. James,et al.  An electrochemical detector array to study cell biology on the nanoscale. , 2002 .

[4]  Maruf Hossain,et al.  Preferential cell attachment to nitrogen-doped diamond-like carbon (DLC:N) for the measurement of quantal exocytosis. , 2009, Biomaterials.

[5]  A. De Luca,et al.  Chronic hypoxia up‐regulates α1H T‐type channels and low‐threshold catecholamine secretion in rat chromaffin cells , 2007, The Journal of physiology.

[6]  Shubhra Gangopadhyay,et al.  Magnetron sputtered diamond-like carbon microelectrodes for on-chip measurement of quantal catecholamine release from cells , 2008, Biomedical microdevices.

[7]  J. Luong,et al.  Boron-doped diamond electrode: synthesis, characterization, functionalization and analytical applications. , 2009, The Analyst.

[8]  G. Swain,et al.  Characterization and Electrochemical Responsiveness of Boron-Doped Nanocrystalline Diamond Thin-Film Electrodes , 2003 .

[9]  E. Neher,et al.  Protein Kinase C Enhances Exocytosis from Chromaffin Cells by Increasing the Size of the Readily Releasable Pool of Secretory Granules , 1996, Neuron.

[10]  Yanli Zhou,et al.  The application of boron-doped diamond electrodes in amperometric biosensors. , 2009, Talanta.

[11]  Paolo Ariano,et al.  A diamond-based biosensor for the recording of neuronal activity. , 2009, Biosensors & bioelectronics.

[12]  R. Tsien,et al.  Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores. , 1989, The Journal of biological chemistry.

[13]  D. Vandael,et al.  PDE type-4 inhibition increases L-type Ca2+ currents, action potential firing, and quantal size of exocytosis in mouse chromaffin cells , 2009, Pflügers Archiv - European Journal of Physiology.

[14]  J. Castracane,et al.  Amperometric detection of quantal catecholamine secretion from individual cells on micromachined silicon chips. , 2003, Analytical chemistry.

[15]  Fwu-Shan Sheu,et al.  Microelectrode array biochip: tool for in vitro drug screening based on the detection of a drug effect on dopamine release from PC12 cells. , 2006, Analytical chemistry.

[16]  W. Ebert,et al.  Bias enhanced nucleation of diamond on silicon (100) in a HFCVD system , 2003 .

[17]  K. Gillis,et al.  On-chip amperometric measurement of quantal catecholamine release using transparent indium tin oxide electrodes. , 2006, Analytical chemistry.

[18]  Robert H. Chow,et al.  Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells , 1992, Nature.

[19]  V. Parpura.  Nanofabricated carbon-based detector. , 2005, Analytical Chemistry.

[20]  G. M. Swain,et al.  Electrochemical monitoring of nitric oxide released by myenteric neurons of the guinea pig ileum , 2008, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.