利用介電濕潤與絕緣結構介電泳技術製作新型細胞膜片電位箝制晶片

Abstract

細胞膜片電位箝制法(patch clamp technique)是目前研究細胞膜上離子通道蛋白質微小電流變化的標準技術，近年來透過半導體製程的方式可將細胞膜片電位箝制法直接製作在晶片上(patch clamp chip)，除了可避免人為操作上的困難，還可以製作陣列型的箝制晶片(multi-patch)。而我們的研究主要是設計一個新型細胞膜片電位箝制晶片，希望透過介電濕潤技術與絕緣結構介電泳技術來操控脂質體以達到自動化形成脂質雙層膜並進行細胞膜蛋白的電流檢測。目前我們已成功地利用共平面電極介電濕潤的設計來驅動氯化鉀電解液，並進行液滴的切割、合併。另一方面以DNA做為測試絕緣結構介電泳的樣本，分別針對平面與立體絕緣結構介電泳進行模擬與實驗分析。而脂質實驗的部分我們則是在表面修飾過後的晶片上製作出人造平面脂質雙層膜，並嘗試利用微壓印技術與電生成脂質體技術來製作固定大小的脂質體，希望未來可以直接操控脂質體自動形成平面雙層膜以供離子通道電流檢測。

The patch clamp technique is a fundamental technique in modern electrophysiology. It can record electrical currents of single ion channel by using a tiny glass pipette to clamp native biomembrane and membrane protein. Recently this technique has been integrated into microfabricated devices, called “patch clamp chips”. Requiring no operators, patch clamp chips provide automatic and precise manipulation. In this research, we propose a novel patch clamp chip by using electrowetting-on-dielectric (EWOD) and insulator-based dielectrophoresis (iDEP) techniques to manipulate bio-fluids, bio-objects, and liposome. EWOD has been successfully used to transport, cut, and merge potassium chloride droplets in single-sided electrode devices. 2D and 3D iDEP have been demonstrated to trap DNA molecules. Microcontact printing and electroformation were investigated to prepare monodispersed giant liposomes, although not very successfully. We used painted methods to form artificial lipid bilayer and observed it under a microscope. Several basic techniques were examined in this research to fulfill an automatic patch clamp chip for membrane protein study. We hope to demonstrate a high-throughput drug screening microdevice in the future on the basis of integration of the techniques investigated in this research.