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Lateral micro fluidic channels array chip fabrication for automated patch clamp application

机译:横向微流体通道阵列芯片制造,用于自动膜片钳应用

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Traditionally, patch-clamp recording is accomplished with a micromanipulator-positioned glass pipette under a microscope. A cell membrane patch is sucked into the glass pipette and forms a high electrical resistance seal. The high cost and labor-intensive methods of conventional patch clamp have prevented the full potential of ion channels as a drug target class being fully realized. Automated patch clamp systems have recently been developed, in order to inexpensively collect large amounts of data in a shorter period of time. More common automation patch-clamp systems use microchips with tiny (1–2μm) holes in a plate instead of pipettes to create the gigaseals and record from single cells. In our previously reported works, a lateral aperture of a buried micro channel was demonstrated, which differs from the common planar patch aperture and is easier fluidic integration and packaging, higher-density array comparing with planar patch aperture. In this paper, we present the optimized fabrication process and integrated the optimized fabrication process to a new designed lateral patch-clamp array chip with 12 independent lateral patch-clamping sites for patch clamp application. At last, the new designed lateral patch clamp devices are utilized to conduct whole cell patch clamp measurements in rat insulinoma (INS-1) cells. High gigaseals (>1 GΩ) were formed between the glass capillary apertures and INS-1 cells. Steady state I-V plots elicited characteristic ion channel properties and longevity of the whole cell mode could be maintained for 1 h without any breakage of the gigaseals, which long enough to apply various compounds and ion channel drugs.
机译:传统上,膜片钳记录是在显微镜下用放置有微型操纵器的玻璃移液器完成的。细胞膜贴剂被吸到玻璃移液管中并形成高电阻密封。常规膜片钳的高成本和劳动密集型方法阻止了离子通道作为药物目标类别的全部潜力得到充分实现。最近开发了自动膜片钳系统,以便在较短的时间内廉价地收集大量数据。更常见的自动化膜片钳系统使用的是微孔板(而不是移液管)中的微孔板(1-2μm),而不是用移液管来创建千兆字节并从单个细胞中进行记录。在我们先前报道的工作中,展示了掩埋微通道的侧向孔径,该孔径不同于常见的平面贴片孔径,并且与平面贴片孔径相比,更易于流体集成和包装,密度更高。在本文中,我们介绍了优化的制造工艺,并将优化的制造工艺集成到了新设计的横向贴片钳阵列芯片中,该芯片具有12个独立的横向贴片钳位点,用于贴片钳应用。最后,新设计的横向膜片钳设备被用于在大鼠胰岛素瘤(INS-1)细胞中进行全细胞膜片钳测量。在玻璃毛细管孔和INS-1细胞之间形成了较高的千兆凝胶(> 1GΩ)。稳态I-V图可得出特征性的离子通道特性,并且整个细胞模式的寿命可以维持1 h,而不会损坏gigaseal,这足以应用各种化合物和离子通道药物。

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