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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Microelectrochemical cell arrays for whole-cell currents recording through ion channel proteins based on trans-electroporation approach
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Microelectrochemical cell arrays for whole-cell currents recording through ion channel proteins based on trans-electroporation approach

机译:基于跨电电穿孔方法通过离子通道蛋白记录的全电池电流的微电色细胞阵列

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摘要

High electrostability and long life-time of planar chip technology are crucial for electrophysiological measurements such as ionic current recording through ion channel proteins embedded in biological cell membrane. In this paper, we propose a novel planar microchip integrated with microelectrochemical cell array toward to a feasible solution for ion channel screening with high resolution and long life-time. In order to reduce the interference from the leakage currents, a synthetic lipid bilayer is applied to form a high sealing resistance. The whole-cell electrical access can be constructed via electroporating the lipid bilayer in close proximity to the cell membrane. Parameters of electroporation including amplitude and time scale are firstly optimized by using parallel electroporation to the lipid bilayers. In this approach, individual cells can be trapped to the target positions by applying dielectrophoresis (DEP) manipulation. Poly (ethylene glycol) (PEG) is employed with low concentration to facilitate the closed contact between the cells and the lipid bilayer to increase the efficiency of the whole-cell mode construction. Through this chip based method, stable current recordings through inward rectifier potassium (Kir) ion channels embedded in rat basophilic leukemia (RBL-1) cell membrane are achieved with high electrical sealing resistance (over 1 GΩ). In addition, without need for complex fluidic connections, this method allows for an easy operation and further miniaturization of the measuring system.
机译:平面芯片技术的高静电性和长寿命对于电生理测量至关重要,例如通过在生物细胞膜中嵌入的离子通道蛋白质的离子电流记录。在本文中,我们提出了一种与微电子细胞阵列集成的新型平面微芯片,朝向具有高分辨率和长寿命的离子通道筛选的可行解决方案。为了减少漏电流的干扰,施加合成脂质双层以形成高密封性。通过将脂质双层电穿孔靠近细胞膜,可以通过将整个细胞电入接近构成。通过使用平行电穿孔到脂质双层,首先优化包括幅度和时间尺度的电穿孔的参数。在这种方法中,通过施加介电泳(DEP)操纵,可以通过施加介质来捕获各个细胞。聚(乙二醇)(PEG)用低浓度,以促进细胞和脂质双层之间的闭合接触,以提高全细胞模式结构的效率。通过该芯片的方法,通过向内浸没在大鼠嗜碱性白血病(RBL-1)细胞膜中通过向内整流钾(KIR)离子通道的稳定电流记录通过高电密封电阻(超过1gΩ)来实现。另外,不需要复杂的流体连接,该方法允许易于操作和进一步的测量系统小型化。

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