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Hourglass SiO2 coating increases the performance of planar patch-clamp

Paper ID Volume ID Publish Year Pages File Format Full-Text
25245 43562 2006 13 PDF Available
Title
Hourglass SiO2 coating increases the performance of planar patch-clamp
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 SiO2 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 SiO2 led to an hourglass structure of the pore and a tighter seal than that offered by a flat, thermal SiO2 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.

Keywords
Planar patch-clamp; BK(Ca) channels; Inducible expression system; IRK1 channels; PECVD coating; AFM; SEM
First Page Preview
Hourglass SiO2 coating increases the performance of planar patch-clamp
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 125, Issue 1, 20 August 2006, Pages 142–154
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering