Nanopore-based technologies are highly adaptable supports for developing label-free sensor chips to characterize lipid bilayers, membrane proteins, and nucleotides. We utilized a single nanopore chip to study the electrophysiology of the epithelial Na⁺ channel (ENaC) incorporated in supported lipid membrane (SLM). An isolated nanopore was developed inside the silicon cavity followed by fusing large unilamellar vesicles (LUVs) of DPPS (1,2-dipalmitoyl-sn-glycero-3-phosphoserine) and DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine) to produce a solvent-free SLM with giga-ohm (GΩ) sealed impedance. The presence and thickness of SLM on the nanopore chip were confirmed using atomic force spectroscopy. The functionality of SLM with and without ENaC was verified in terms of electrical impedance and capacitance by sweeping the frequency from 0.01 Hz to 100 kHz using electrochemical impedance spectroscopy. The nanopore chip exhibits long-term stability for the lipid bilayer before (144 h) and after (16 h) incorporation of ENaC. Amiloride, an inhibitor of ENaC, was utilized at different concentrations to test the integrity of fused ENaC in the lipid bilayer supported on a single nanopore chip. The developed model presents excellent electrical properties and improved mechanical stability of SLM, making this technology a reliable platform to study ion channel electrophysiology.

中文翻译：

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使用单个纳米孔芯片嵌入支持脂质双层中的上皮钠通道（ENaC）的电生理

基于纳米孔的技术是开发无标记传感器芯片以表征脂质双层，膜蛋白和核苷酸的高度适应性支持。我们利用单个纳米孔芯片来研究支持脂质膜（SLM）中整合的上皮Na ⁺通道（ENaC）的电生理。在硅腔内形成了一个孤立的纳米孔，然后将DPPS（1,2-二棕榈酰-sn-甘油-3-磷酸丝氨酸）和DPPE（1,2-二棕榈酰-sn）的大单层囊泡（LUV）融合在一起-glycero-3-phosphoethanolamine）生成具有千兆欧（GΩ）密封阻抗的无溶剂SLM。使用原子力光谱法确认了纳米孔芯片上SLM的存在和厚度。通过使用电化学阻抗谱将频率从0.01 Hz扫描到100 kHz，可以在电阻抗和电容方面验证具有和不具有ENaC的SLM的功能。纳米孔芯片在掺入ENaC之前（144 h）和之后（16 h）对脂质双层表现出长期稳定性。阿米洛利（ENaC的抑制剂）以不同的浓度使用，以测试在单个纳米孔芯片上支撑的脂质双层中融合的ENaC的完整性。开发的模型具有出色的电性能，并改善了SLM的机械稳定性，