Characterization of a patch-clamp microchannel array towards neuronal networks analysis

The attempt to combine the planar patch clamping idea with the microelectrode array (MEA) concept has led to the fabrication of a patch clamp microchannel array (PCμCA). Such a system is thought to be a powerful framework for neuroscience research and drug screening, as a novel tool for simultaneous patch clamping of cultured cells or neurons in the same network. A disposable silicon/silicon dioxide (Si/SiO2) chip with a microhole array was integrated in a microfluidic system for cell handling, perfusion and electrical recording. Fluidic characterization showed that our PCμCA can work as a precise local perfusion system for chemicals or drugs. Electrical characterization for microholes of 2 μm and 3 μm revealed an access resistance of 8.09 ± 0.84 MΩ and 3.18 ± 0.63 MΩ, respectively. The capacitance was 98.6 ± 13.2 pF. The values are close to what can be expected from theory, but the capacitance is still too high for high resolution recording. The system was tested on HeLa cells: successful cell trapping with a sealing of 40 MΩ was recorded. Modification of the Si/SiO2 chip is needed in order to achieve a better sealing and long-term cell culturing in the PCμCA remains to be tested.

[1]  Diana Conte Camerino,et al.  Ion channel pharmacology , 2007, Neurotherapeutics.

[2]  K. L. Perkins,et al.  Cell-attached voltage-clamp and current-clamp recording and stimulation techniques in brain slices , 2006, Journal of Neuroscience Methods.

[3]  Niels Fertig,et al.  Planar Patch Clamping , 2007 .

[4]  G. Gross,et al.  The use of neuronal networks on multielectrode arrays as biosensors. , 1995, Biosensors & bioelectronics.

[5]  Y. Horio,et al.  Ion channels and diseases , 2002, Medical Electron Microscopy.

[6]  Luke P. Lee,et al.  Open-access microfluidic patch-clamp array with raised lateral cell trapping sites. , 2006, Lab on a chip.

[7]  Wolfgang Walz,et al.  Patch-Clamp Analysis , 2002, Neuromethods.

[8]  Christophe Py,et al.  Cell placement and guidance on substrates for neurochip interfaces , 2010, Biotechnology and bioengineering.

[9]  R A Levis,et al.  Low-noise patch-clamp techniques. , 1998, Methods in enzymology.

[10]  Characterization of a MEMS BioChip for planar patch-clamp recording , 2004 .

[11]  H. Robinson,et al.  Simultaneous induction of pathway-specific potentiation and depression in networks of cortical neurons. , 1999, Biophysical journal.

[12]  C. Cotman,et al.  A microfluidic culture platform for CNS axonal injury, regeneration and transport , 2005, Nature Methods.

[13]  H. Andersson,et al.  Microfluidic devices for cellomics: a review , 2003 .

[14]  R. Peri,et al.  High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology , 2008, Nature Reviews Drug Discovery.

[15]  P. Connolly,et al.  Advantages of using microfabricated extracellular electrodes for in vitro neuronal recording , 1995, Journal of neuroscience research.

[16]  Chang-Yu Chen,et al.  Hourglass-shaped aperture for cellular electrophysiological study , 2007 .

[17]  Conrad D. James,et al.  Patterning Axonal Guidance Molecules Using a Novel Strategy for Microcontact Printing , 2003, Neurochemical Research.

[18]  W. Almers,et al.  Patch voltage clamping with low-resistance seals: loose patch clamp. , 1992, Methods in enzymology.

[19]  Alfred Stett,et al.  CYTOCENTERING: a novel technique enabling automated cell-by-cell patch clamping with the CYTOPATCH chip. , 2003, Receptors & channels.

[20]  Christophe Py,et al.  Neurogenesis and neuronal communication on micropatterned neurochips. , 2005, Biotechnology and bioengineering.

[21]  B. Eversmann,et al.  A 128 × 128 CMOS bio-sensor array for extracellular recording of neural activity , 2003 .

[22]  Growth and synapse formation by identified leech neurones in culture: a review. , 1989, Quarterly journal of experimental physiology.

[23]  Theodore Leng,et al.  The Artificial Synapse Chip: a flexible retinal interface based on directed retinal cell growth and neurotransmitter stimulation. , 2003, Artificial organs.

[24]  C. Py,et al.  Application of polymer microstructures with controlled surface chemistries as a platform for creating and interfacing with synthetic neural networks , 2005, Proceedings. 2005 IEEE International Joint Conference on Neural Networks, 2005..

[25]  Michel De Waard,et al.  Hourglass SiO2 coating increases the performance of planar patch-clamp. , 2006, Journal of biotechnology.

[26]  Xiaobo Wang,et al.  Automated electrophysiology: high throughput of art. , 2003, Assay and drug development technologies.

[27]  J. Judy,et al.  Design and fabrication of a micromachined planar patch-clamp substrate with integrated microfluidics for single-cell measurements , 2006, Journal of Microelectromechanical Systems.

[28]  L. Yobas,et al.  Microfluidic integration of substantially round glass capillaries for lateral patch clamping on chip. , 2007, Lab on a chip.

[29]  A. Offenhäusser,et al.  Patterning chemical stimulation of reconstructed neuronal networks. , 2006, Analytica chimica acta.

[30]  F. Sigworth,et al.  Patch clamp on a chip. , 2002, Biophysical journal.

[31]  D. Schmitt-Landsiedel,et al.  A 128 /spl times/ 128 CMOS bio-sensor array for extracellular recording of neural activity , 2003, 2003 IEEE International Solid-State Circuits Conference, 2003. Digest of Technical Papers. ISSCC..

[32]  Fred J. Sigworth,et al.  An air-molding technique for fabricating PDMS planar patch-clamp electrodes , 2004, Pflügers Archiv.

[33]  M. Alexander,et al.  Principles of Neural Science , 1981 .

[34]  B Sakmann,et al.  Patch clamp techniques for studying ionic channels in excitable membranes. , 1984, Annual review of physiology.

[35]  Thomas M Pearce,et al.  Microtechnology: meet neurobiology. , 2007, Lab on a chip.

[36]  Chang-Yu Chen,et al.  Patch clamping on plane glass-fabrication of hourglass aperture and high-yield ion channel recording. , 2009, Lab on a chip.

[37]  T. Jentsch,et al.  Ion channel diseases. , 2002, Human molecular genetics.

[38]  A Brüggemann,et al.  High quality ion channel analysis on a chip with the NPC technology. , 2003, Assay and drug development technologies.

[39]  B. Wheeler,et al.  Recording from the Aplysia Abdominal Ganglion with a Planar Microelectrode Array , 1986, IEEE Transactions on Biomedical Engineering.

[40]  C. Pudda,et al.  A Silicon-Based "Multi Patch" Device for Ion Channel Current Sensing , 2004 .

[41]  Fei Su,et al.  Microfluidics-Based Biochips: Technology Issues, Implementation Platforms, and Design-Automation Challenges , 2006, IEEE Transactions on Computer-Aided Design of Integrated Circuits and Systems.

[42]  H. Lester,et al.  Ion channel diseases of the central nervous system. , 2006, CNS drug reviews.

[43]  N. Fertig,et al.  Activity of single ion channel proteins detected with a planar microstructure , 2002 .

[44]  B. Sakmann,et al.  Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches , 1981, Pflügers Archiv.

[45]  B. Sakmann,et al.  Single-channel currents recorded from membrane of denervated frog muscle fibres , 1976, Nature.

[46]  W. Nisch,et al.  Patch-clamping of primary cardiac cells with micro-openings in polyimide films , 2003, Medical and Biological Engineering and Computing.

[47]  Martin A. M. Gijs,et al.  Glass reflow on 3-dimensional micro-apertures for electrophysiological measurements on-chip , 2006 .

[48]  S. Cannon Physiologic principles underlying ion channelopathies , 2007, Neurotherapeutics.

[49]  Daniel A. Wagenaar,et al.  The Neurally Controlled Animat: Biological Brains Acting with Simulated Bodies , 2001, Auton. Robots.