Hourglass SiO2 coating increases the performance of planar patch-clamp

Autor: Nathalie Picollet-D'hahan, Christophe Arnoult, Thomas Sordel, Frédérique Marcel, Fabien Sauter, Stéphanie Garnier-Raveaud, Francois Chatelain, Michel Vivaudou, Michel De Waard, Catherine Pudda
Přispěvatelé: 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, Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF)
Rok vydání: 2006
Předmět:
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
Zdroj: Journal of Biotechnology
Journal of Biotechnology, 2006, 125 (1), pp.142-54. ⟨10.1016/j.jbiotec.2006.02.008⟩
Journal of Biotechnology, Elsevier, 2006, 125 (1), pp.142-54. ⟨10.1016/j.jbiotec.2006.02.008⟩
ISSN: 0168-1656
DOI: 10.1016/j.jbiotec.2006.02.008
Popis: 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.
Databáze: OpenAIRE
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