151. Hourglass SiO2 coating increases the performance of planar patch-clamp
- Author
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Nathalie Picollet-D'hahan, Christophe Arnoult, Thomas Sordel, Frédérique Marcel, Fabien Sauter, Stéphanie Garnier-Raveaud, Francois Chatelain, Michel Vivaudou, Michel De Waard, Catherine Pudda, Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire Composants intégrés pour le vivant (LCIV), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Canaux calciques , fonctions et pathologies, Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de biophysique moléculaire et cellulaire (LBMC), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'Energie Atomique, Collaboration, Canepari, Marco, and Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF)
- Subjects
Patch-Clamp Techniques ,[SDV.BIO]Life Sciences [q-bio]/Biotechnology ,MESH: Cricetinae ,02 engineering and technology ,Chemical vapor deposition ,Microscopy, Atomic Force ,Applied Microbiology and Biotechnology ,Ion Channels ,Membrane Potentials ,Planar ,MESH: Cricetulus ,Coating ,Plasma-enhanced chemical vapor deposition ,Cricetinae ,Surface roughness ,MESH: Animals ,PECVD coating ,[INFO.INFO-BT]Computer Science [cs]/Biotechnology ,MESH: Microscopy, Atomic Force ,0303 health sciences ,SEM ,Pipette ,General Medicine ,Silicon Dioxide ,021001 nanoscience & nanotechnology ,MESH: Reproducibility of Results ,Optoelectronics ,AFM ,0210 nano-technology ,Plasmids ,Biotechnology ,Materials science ,MESH: Microscopy, Electron, Scanning ,Bioengineering ,Nanotechnology ,CHO Cells ,engineering.material ,Transfection ,Capacitance ,MESH: Silicon Dioxide ,Cell Line ,03 medical and health sciences ,Cricetulus ,MESH: CHO Cells ,MESH: Plasmids ,MESH: Patch-Clamp Techniques ,Animals ,Humans ,MESH: Membrane Potentials ,[SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials ,030304 developmental biology ,MESH: Humans ,business.industry ,MESH: Transfection ,Reproducibility of Results ,MESH: Cell Line ,[SDV.BIO] Life Sciences [q-bio]/Biotechnology ,[SDV.IB.BIO] Life Sciences [q-bio]/Bioengineering/Biomaterials ,Surface coating ,planar patch-clamp ,[INFO.INFO-BT] Computer Science [cs]/Biotechnology ,BK(Ca) channels ,MESH: Ion Channels ,Microscopy, Electron, Scanning ,engineering ,IRK1 channels ,business - Abstract
International audience; 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.
- Published
- 2006