Indium Tin Oxide devices for amperometric detection of vesicular release by single cells.

The microfabrication and successful testing of a series of three ITO (Indium Tin Oxide) microsystems for amperometric detection of cells exocytosis are reported. These microdevices have been optimized in order to simultaneously (i) enhance signal-to-noise ratios, as required electrochemical monitoring, by defining appropriate electrodes geometry and size, and (ii) provide surface conditions which allow cells to be cultured over during one or two days, through apposite deposition of a collagen film. The intrinsic electrochemical quality of the microdevices as well as the effect of different collagen treatments were assessed by investigating the voltammetric responses of two classical redox systems, Ru(NH(3))(6)(3+/2+) and Fe(CN)(6)(3-/4-). This established that a moderate collagen treatment does not incur any significant alteration of voltammetric responses or degradation of the excellent signal-to-noise ratio. Among these three microdevices, the most versatile one involved a configuration in which the ITO microelectrodes were delimited by a microchannel coiled into a spiral. Though providing extremely good electrochemical responses this specific design allowed proper seeding and culture of cells permitting either single cell or cell cluster stimulation and analysis.

[1]  C. Guillén,et al.  Comparison study of ITO thin films deposited by sputtering at room temperature onto polymer and glass substrates , 2005 .

[2]  Eunkyoung Kim,et al.  Tetrazine-based electrofluorochromic windows: Modulation of the fluorescence through applied potential , 2009 .

[3]  Alan Morgan,et al.  Secretory granule exocytosis. , 2003, Physiological reviews.

[4]  H. Chan,et al.  Release monitoring of single cells on a microfluidic device coupled with fluorescence microscopy and electrochemistry. , 2010, Biomicrofluidics.

[5]  Yu Wang,et al.  Monitoring of dopamine release in single cell using ultrasensitive ITO microsensors modified with carbon nanotubes. , 2011, Biosensors & bioelectronics.

[6]  Maruf Hossain,et al.  Controlled on-chip stimulation of quantal catecholamine release from chromaffin cells using photolysis of caged Ca2+ on transparent indium-tin-oxide microchip electrodes. , 2008, Lab on a chip.

[7]  Wei Wang,et al.  Monitoring of vesicular exocytosis from single cells using micrometer and nanometer-sized electrochemical sensors , 2009, Analytical and bioanalytical chemistry.

[8]  Y. Tu,et al.  Studies on the electrochemiluminescent behavior of luminol on indium tin oxide (ITO) glass , 2010 .

[9]  Fwu-Shan Sheu,et al.  In situ temporal detection of dopamine exocytosis from L-dopa-incubated MN9D cells using microelectrode array-integrated biochip , 2006 .

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

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

[12]  Khajak Berberian,et al.  Electrochemical imaging of fusion pore openings by electrochemical detector arrays. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Christian Amatore,et al.  Electrochemical monitoring of single cell secretion: vesicular exocytosis and oxidative stress. , 2008, Chemical reviews.

[14]  A. Pasquarelli,et al.  Diamond microelectrodes arrays for the detection of secretory cell activity , 2011 .

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

[16]  Yong Chen,et al.  Coupling of electrochemistry and fluorescence microscopy at indium tin oxide microelectrodes for the analysis of single exocytotic events. , 2006, Angewandte Chemie.

[17]  J. Savéant,et al.  Charge transfer at partially blocked surfaces , 1983 .

[18]  Kevin D. Gillis,et al.  Microwell device for targeting single cells to electrochemical microelectrodes for high-throughput amperometric detection of quantal exocytosis. , 2011, Analytical chemistry.

[19]  E. Karatekin,et al.  Coupling amperometry and total internal reflection fluorescence microscopy at ITO surfaces for monitoring exocytosis of single vesicles. , 2011, Angewandte Chemie.

[20]  Matthieu Piel,et al.  Microfluidic tools for cell biological research. , 2010, Nano today.

[21]  Gregory M Dittami,et al.  Electrically evoking and electrochemically resolving quantal release on a microchip. , 2010, Lab on a chip.

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

[23]  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.

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

[25]  Maruf Hossain,et al.  Automated targeting of cells to electrochemical electrodes using a surface chemistry approach for the measurement of quantal exocytosis. , 2010, ACS chemical neuroscience.

[26]  R. Wightman,et al.  Spatio-temporal resolution of exocytosis from individual cells. , 1998, Annual review of biophysics and biomolecular structure.

[27]  Carlos M. Fernández-Peruchena,et al.  Fusion pore regulation of transmitter release , 2005, Brain Research Reviews.

[28]  Peng Chen,et al.  Micro- and nanotechnologies for study of cell secretion. , 2011, Analytical chemistry.

[29]  K. Gillis,et al.  A microfluidic cell trap device for automated measurement of quantal catecholamine release from cells. , 2009, Lab on a chip.

[30]  R. Kennedy,et al.  Effects of Intravesicular H+ and Extracellular H+ and Zn2+ on Insulin Secretion in Pancreatic Beta Cells* , 1997, The Journal of Biological Chemistry.

[31]  Christy L Haynes,et al.  Bioanalytical tools for single-cell study of exocytosis , 2010, Analytical and bioanalytical chemistry.

[32]  Christopher M.A. Brett,et al.  Electrochemistry: Principles, Methods, and Applications , 1993 .

[33]  Arto Heiskanen,et al.  Chip Based Electroanalytical Systems for Cell Analysis , 2008 .

[34]  Weishan Li,et al.  DNA-enhanced assembly of [Ru(bpy)2ITATP]3+/2+ on an ITO electrode , 2007 .

[35]  Lisa J. Mellander,et al.  Electrochemical probes for detection and analysis of exocytosis and vesicles. , 2010, Chemphyschem : a European journal of chemical physics and physical chemistry.

[36]  R. Wightman Probing Cellular Chemistry in Biological Systems with Microelectrodes , 2006, Science.

[37]  C. Amatore,et al.  Comparison of apex and bottom secretion efficiency at chromaffin cells as measured by amperometry. , 2007, Biophysical chemistry.

[38]  R. Stark,et al.  Cell proliferation assays on plasma activated SU-8 , 2008 .

[39]  K. Gillis,et al.  Measuring secretion in chromaffin cells using electrophysiological and electrochemical methods , 2007, Acta physiologica.

[40]  Andrew G Ewing,et al.  Analytical approaches to investigate transmitter content and release from single secretory vesicles , 2010, Analytical and bioanalytical chemistry.

[41]  A. Teschemacher Real-time measurements of noradrenaline release in periphery and central nervous system , 2005, Autonomic Neuroscience.

[42]  R. Kennedy,et al.  Amperometric monitoring of chemical secretions from individual pancreatic beta-cells. , 1993, Analytical chemistry.

[43]  A. Pasquarelli,et al.  Nanocrystalline diamond microelectrode arrays fabricated on sapphire technology for high-time resolution of quantal catecholamine secretion from chromaffin cells. , 2010, Biosensors & bioelectronics.

[44]  Allen J. Bard,et al.  Electrochemical Methods: Fundamentals and Applications , 1980 .

[45]  H. Maeda An Atomic Force Microscopy Study of Ordered Molecular Assemblies and Concentric Ring Patterns from Evaporating Droplets of Collagen Solutions , 1999 .