膜蛋白嵌入脂質雙層膜與數位微流體晶片之研究

Abstract

人工的脂質雙層膜被廣泛的運用在膜蛋白的電生理量測及研究上，而近年來對於實驗室晶片的發展，讓建構人工脂質雙層膜的技術擴展到微流體晶片上。本實驗室便成功的以數位微流體晶片技術，發展出以介電濕潤驅動包覆式液珠而形成脂質雙層膜的系統，可在微米等級的孔洞上，形成懸浮式的脂質雙層膜，並以光學觀測和電訊號的量測進行脂質雙層膜成膜確認，並在其後嵌入膜蛋白以電生理訊號進行離子通道的研究。我們成功的以本論文中建構的晶片系統對Gramicidin和□-HL進行電生理量測，更近一步的利用光學螢光觀測V-PPase水解焦磷酸並傳輸質子的生理運動。最後，我們在論文中對於此系統做最佳化，希望能改善其不足之處。我們設計出加壓薄化系統希望能掌控成膜的過程、全油環境希望能讓成膜環境更穩定、平面式的銀/氯化銀電極讓晶片的結構更完整。

Artificial lipid bilayer membrane (BLM) is widely used in the electrophysiological study of membrane proteins. Recently, researchers make effort to introduce the technique of microfluidic into BLM formation to realize bio-assay in a single chip which calls lab-on-a-chip. In this research, we develop a system to form a BLM suspends in a 100 □m2 hole with the manipulation of capsulated droplet by EWOD (electrowetting-on-dielectric) and proceed with optical and electrical analysis to confirm the formation of BLM. After BLM formation, we incorporate membrane protein to study the ion channel by electrophysiology. Here we successfully to employ the BLM formation system we developed in the study of two ion channel peptide, Gramicidin and α-hemolysin. Moreover, the PPi hydrolysis and inducing of proton transportation by V-PPase was studied under a fluorescent microscope. At the end, we propose new designs to optimize this membrane formation system, including pressure-assist membrane formation, on chip planer Ag/AgCl electrode to replace the probe based electrophysiology analysis and oil bath environment to make membrane more stable from the evaporation.