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PMID: 16595156 Published · ppublish English Journal Article

Hourglass SiO2 coating increases the performance of planar patch-clamp.

Journal of biotechnology ·Vol. 125 ·No. 1 ·2006-08-20 ·Pages 142-54

Sordel T, Garnier-Raveaud S, Sauter F, Pudda C, Marcel F, De Waard M, Arnoult C, Vivaudou M, Chatelain F, Picollet-D'hahan N

Abstract

Obtaining high-throughput electrophysiological recordings is an ongoing challenge in ion channel biophysics and drug discovery. One particular area of development is the replacement of glass pipettes with planar devices in order to increase throughput. However, successful patch-clamp recordings depend on a surface coating which ideally should promote and stabilize giga-seal formation. Here, we present data supporting the use of a structured SiO(2) coating to improve the ability of cells to form a "seal" with a planar patch-clamp substrate. The method is based on a correlation study taking into account structure and size of the pores, surface roughness and chip capacitance. The influence of these parameters on the quality of the seal was assessed. Plasma-enhanced chemical vapour deposition (PECVD) of SiO(2) led to an hourglass structure of the pore and a tighter seal than that offered by a flat, thermal SiO(2) surface. The performance of PECVD chips was validated by recording recombinant potassium channels, BK(Ca), expressed in stable HEK-293 cell lines and in inducible CHO cell lines and low conductance IRK1, and endogenous cationic currents from CHO cells. This multiparametric investigation led to the production of improved chips for planar patch-clamp applications which allow electrophysiological recordings from a wide range of cell lines.

MeSH Terms
Animals CHO Cells Cell Line Cricetinae Cricetulus Humans Ion Channels/genetics,physiology Membrane Potentials/physiology Microscopy, Atomic Force Microscopy, Electron, Scanning Patch-Clamp Techniques/instrumentation,methods Plasmids/genetics Reproducibility of Results Silicon Dioxide/chemistry Transfection
Chemicals
Ion Channels Silicon Dioxide
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Sordel Thomas
Laboratoire Biopuces, CEA, 17 Rue des Martyrs, 38054 Grenoble Cedex 9, France.
Garnier-Raveaud Stéphanie
Sauter Fabien
Pudda Catherine
Marcel Frédérique
De Waard Michel
Arnoult Christophe
Vivaudou Michel
Chatelain François
Picollet-D'hahan Nathalie
Article Info
Journal
Journal of biotechnology
Abbr.
J Biotechnol
ISSN
0168-1656
Published
2006-08-20
Epub
2006-00-03
Pages
142-54
Language
English
Region
Netherlands
NLM ID
8411927
Subset
IM
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