以共平面式多電極陣列介電濕潤晶片合成DNA磁球微粒檢測探針之應用

A Multiarray Coplanar-Type-Electrodes EWOD Chip Apply to Magnetic-bead Probes Formation of DNA Detection

蘇琮堯(Tsung-Yao Su)

指導教授 ： 饒達仁

國立清華大學/工學院/奈米工程與微系統研究所/碩士(2010年)

摘要

本研究就主要以介電濕潤（Electrowetting-on-dielectric, EWOD）法作為操控微液滴之平台，藉由共平面式多電極之操控陣列可達到同步處理多個DNA檢體，以油包水之操控方式操控包括不同生醫檢體包含：去氧核醣核酸（DNA）、磁球微粒（Magnetic beads）及緩衝液（Buffer）等，並以此完成一可檢測如DNA單點突變（Single-nucleotide Polymorphism, SNP）之磁球探針。
操控晶片利用ITO（Indium Tin Oxide）玻璃製作出共平面式操控電極，包含多個儲存槽及傳輸電極，搭配無電極之疏水上蓋達到液滴於多個儲存槽同步產生、移動、混合與分離等效果，由於ITO玻璃本身為透明且具有良好之光學性質，因此在後續的光學檢測及影像擷取皆非常便利。透過此共平面式介電濕潤平台的建構，本實驗完成一個開放式的數位微流體系統（Digital Micro Fluidic System），使用者可以更有彈性的進行液珠的傳輸及混合等操控。
本研究將SNP檢測探針會使用之核酸接合流程在晶片上實行，並對含磁球之液滴做操控測試，最後成功於晶片上完成核酸接合檢測及磁球探針合成步驟，利用量子點（QDots）與探針接合可於螢光顯微鏡下確認探針合成成效，達到實驗室晶片操作目標。

關鍵字

介電濕潤 ； 生物探針 ； 磁球標示

並列摘要

This study bases on the technique, “Electrowetting on Dielectric (EWOD),” which is actuated by addressable coplanar electrodes for micro droplets manipulation. Multiple bio-reagents such as DNA, magnetic beads and buffer have manipulated synchronous with different concentration. All of the droplets are driven with water oil core-shell type, which are surrounded by silicon oil. All the droplets are generated homogeneously with ultra microvolume(0.3μL).
A coplanar EWOD chip with several reservoirs and electrodes, is fabricated by indium tin oxide (ITO) glass, which can drive droplets for creating, transporting, merge and splitting. All the fluidics movement is confined between two plates, a bottom plate for the coplanar chip, and another hydrophobic plate with SU8 structure for the top. Detections and capturing of images are easy and convenient through transparent ITO glasses, which are well characteristic of optic. Through the construction of this addressable coplanar-electrode EWOD chip, the study demonstrates a flexible platform for Digital Micro Fluidic (DMF) Systems to manipulate micro droplets.
To perform a bio-application on the EWOD chip, we transfer a DNA ligation process on this device, which is a common procedure for biomedical analysis. Droplets with magnetic beads are also manipulated on the chip. Moreover, a probe formation process of SNP detection is also completed which confirmed by QDots labeling. The result shows that probes synthesized by EWOD chip are better than by hand. Thus, in this paper, we wish the device could be integrated with other systems to achieve the goal of Micro Total Analysis System (μTAS) and Lab on a Chip.

並列關鍵字

electrowetting on dielectric ； EWOD ； bio-probe ； magnetic-bead labeling

參考文獻

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