Xu Jia, Guia António, Rothwarf David, Huang Mingxian, Sithiphong Khachone, Ouang Joe, Tao Guoliang, Wang Xiaobo, Wu Lei
AVIVA Biosciences Corp, San Diego, CA 92121, USA.
Assay Drug Dev Technol. 2003 Oct;1(5):675-84. doi: 10.1089/154065803770381039.
Although conventional patch-clamp methods provide high information content, they are labor-intensive and suffer from low throughput and high overall cost. Several approaches for achieving high throughput electrophysiology are under development, among which microchip-based patch-clamp systems uniquely achieve a higher degree of miniaturization, faster perfusion and mixing, and lower reagent cost without losing information content. The goal of this study was to establish a benchmark for our biochip technology with 52 chips tested sequentially. We demonstrate that our microfabrication and processing technology is sufficiently mature to produce a consistent hole size. We further demonstrate high-quality planar whole-cell patch clamping with >75% overall success rate at achieving gigaohm seals, followed by stable whole-cell access lasting at least 15 min with access resistance (Ra) below 15 MOmega and membrane resistance (Rm) above 200 MOmega. These biochips are ideally suited for high throughput compound screening for ion channel targets.
尽管传统的膜片钳方法能提供丰富的信息，但它们劳动强度大，通量低且总成本高。目前正在开发几种实现高通量电生理学的方法，其中基于微芯片的膜片钳系统在不失信息含量的情况下，独特地实现了更高程度的小型化、更快的灌注和混合以及更低的试剂成本。本研究的目标是通过对52个芯片进行顺序测试，为我们的生物芯片技术建立一个基准。我们证明，我们的微加工和处理技术已经足够成熟，能够产生一致的孔尺寸。我们进一步展示了高质量的平面全细胞膜片钳记录，实现千兆欧封接的总体成功率超过75%，随后是稳定的全细胞通路，持续至少15分钟，通路电阻（Ra）低于15兆欧，膜电阻（Rm）高于200兆欧。这些生物芯片非常适合用于离子通道靶点的高通量化合物筛选。
