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Mammalian electrophysiology on a microfluidic platform

机译:微流控平台上的哺乳动物电生理

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摘要

The recent development of automated patch clamp technology has increased the throughput of electrophysiology but at the expense of visual access to the cells being studied. To improve visualization and the control of cell position, we have developed a simple alternative patch clamp technique based on microfluidic junctions between a main chamber and lateral recording capillaries, all fabricated by micromolding of polydimethylsiloxane (PDMS). PDMS substrates eliminate the need for vibration isolation and allow direct cell visualization and manipulation using standard microscopy. Microfluidic integration allows recording capillaries to be arrayed 20 mu m apart, for a total chamber volume of < 0.5 nl. The geometry of the recording capillaries permits high-quality, stable, whole-cell seals despite the hydrophobicity of the PDMS surface. Using this device, we are able to demonstrate reliable whole-cell recording of mammalian cells on an inexpensive microfluidic platform. Recordings of activation of the voltage-sensitive potassium channel Kv2.1 in mammalian cells compare well with traditional pipette recordings. The results make possible the integration of whole-cell electrophysiology with easily manufactured microfluidic lab-on-a-chip devices.
机译:自动化膜片钳技术的最新发展增加了电生理学的通量,但以可视方式访问正在研究的细胞为代价。为了改善可视化和细胞位置的控制,我们已经开发了一种简单的替代膜片钳技术,该技术基于主腔室和侧向记录毛细管之间的微流体连接,均由聚二甲基硅氧烷(PDMS)微模制而成。 PDMS基板消除了隔振的需要,并允许使用标准显微镜直接观察和处理细胞。微流控集成允许记录毛细管以20μm的间隔排列,总腔室容积小于0.5 nl。尽管PDMS表面具有疏水性,但记录毛细管的几何形状仍可实现高质量,稳定的全细胞密封。使用此设备,我们能够在便宜的微流控平台上证明哺乳动物细胞的可靠全细胞记录。与传统的移液器记录相比,哺乳动物细胞中压敏钾通道Kv2.1的激活记录很好。结果使全细胞电生理学与易于制造的微流控芯片实验室设备的集成成为可能。

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