Chen Chang-Yu, Tu Ting-Yuan, Chen Chang-Hung, Jong De-Shien, Wo Andrew M
Institute of Applied Mechanics, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei, 106, Taiwan.
Lab Chip. 2009 Aug 21;9(16):2370-80. doi: 10.1039/b901025d. Epub 2009 May 14.
Planar patch-clamp has revolutionized ion-channel measurement by eliminating laborious manipulation from the traditional micropipette approach and enabling high throughput. However, low yield in gigaseal formation and/or relatively high cost due to microfabricated processes are two main drawbacks. This paper presents patch clamping on glass substrate-an economical solution without sacrificing gigaseal yield rate. Two-stage CO(2) laser drilling methodology was used to generate an hourglass, funnel-like aperture of a specified diameter with smooth and debris-free surfaces on 150 microm borosilicate cover glass. For 1-3 microm apertures as patch-clamp chips, seal resistance was tested on human embryonic kidney, Chinese hamster ovary, and Jurkat T lymphoma cells with a gigaseal success rate of 62.5%, 43.6% and 66.7% respectively. Results also demonstrated both whole-cell and single channel recording on endogenously expressed ion channels to confirm the capability of different patch configurations.
平面膜片钳技术通过摒弃传统微吸管方法中繁琐的操作并实现高通量，彻底改变了离子通道测量技术。然而，千兆欧姆封接形成率低和/或由于微加工工艺导致成本相对较高是两个主要缺点。本文介绍了在玻璃基板上进行膜片钳操作——一种在不牺牲千兆欧姆封接率的情况下的经济解决方案。采用两级二氧化碳激光钻孔方法，在150微米的硼硅酸盐盖玻片上生成了具有指定直径的沙漏形、漏斗状孔径，表面光滑且无碎屑。对于作为膜片钳芯片的1 - 3微米孔径，在人胚肾细胞、中国仓鼠卵巢细胞和Jurkat T淋巴瘤细胞上测试了封接电阻，千兆欧姆封接成功率分别为62.5%、43.6%和66.7%。结果还证明了在内源表达的离子通道上进行全细胞和单通道记录，以确认不同膜片配置的能力。
