Ion channel recordings on an injection-molded polymer chip

被引:13
|
作者
Tanzi, Simone [1 ]
Matteucci, Marco [1 ]
Christiansen, Thomas Lehrmann [1 ]
Friis, Soren [2 ]
Christensen, Mette Thylstrup [2 ]
Garnaes, Joergen [3 ]
Wilson, Sandra [2 ]
Kutchinsky, Jonatan [2 ]
Taboryski, Rafael [1 ]
机构
[1] Tech Univ Denmark, Dept Micro & Nanotechnol, DK-2800 Lyngby, Denmark
[2] Soph Biosci AS, DK-2750 Ballerup, Denmark
[3] Danish Fundamental Metrol, Matemat Torvet, DK-2800 Lyngby, Denmark
关键词
PATCH-CLAMP TECHNOLOGY; DRUG DISCOVERY; ELECTROOSMOTIC MOBILITY; HYDROPHOBIC POLYMERS; MICROFLUIDIC SYSTEM; ASSAY TECHNOLOGIES; WATER-STRUCTURE; SODIUM-CHANNEL; ELECTROPHYSIOLOGY; MEMBRANE;
D O I
10.1039/c3lc50760b
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In this paper, we demonstrate recordings of the ion channel activity across the cell membrane in a biological cell by employing the so-called patch clamping technique on an injection-molded polymer microfluidic device. The findings will allow direct recordings of ion channel activity to be made using the cheapest materials and production platform to date and with the potential for very high throughput. The employment of cornered apertures for cell capture allowed the fabrication of devices without through holes and via a scheme comprising master origination by dry etching in a silicon substrate, electroplating in nickel and injection molding of the final part. The most critical device parameters were identified as the length of the patching capillary and the very low surface roughness on the inside of the capillary. The cross-sectional shape of the orifice was found to be less critical, as both rectangular and semicircular profiles seemed to have almost the same ability to form tight seals with cells with negligible leak currents. The devices were functionally tested using human embryonic kidney cells expressing voltage-gated sodium channels (Na(v)1.7) and benchmarked against a commercial state-of-the-art system for automated ion channel recordings. These experiments considered current-voltage (IV) relationships for activation and inactivation of the Na(v)1.7 channels and their sensitivity to a local anesthetic, lidocaine. Both IVs and lidocaine dose-response curves obtained from the injection-molded polymer device were in good agreement with data obtained from the commercial system.
引用
收藏
页码:4784 / 4793
页数:10
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