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系統識別號 U0026-0812200915275667
論文名稱(中文) 整合氣動式常關閥門之毛細管電泳晶片應用於去氧核醣核酸預濃縮及分離之研究
論文名稱(英文) Micro-Fabricated Capillary Electrophoresis Chip for DNA Pre-concentration and Separation Utilizing a Normally-closed Valve
校院名稱 成功大學
系所名稱(中) 工程科學系碩博士班
系所名稱(英) Department of Engineering Science
學年度 97
學期 2
出版年 98
研究生(中文) 郭貞華
研究生(英文) Chen-Hua Kuo
學號 n9696440
學位類別 碩士
語文別 英文
論文頁數 59頁
口試委員 口試委員-饒達仁
口試委員-林哲信
指導教授-李國賓
中文關鍵字 預濃縮  DNA分離  毛細管電泳晶片  常關式閥門 
英文關鍵字 DNA separation  micro capillary electrophoresis  normally-closed valve  pre-concentration 
學科別分類
中文摘要 本論文研發了一個簡單及連續式之毛細管電泳晶片系統，將一個氣動式之常關閥門整合於晶片上，以達成去氧核醣核酸(deoxyribonucleic acid, DNA)樣品之預濃縮以及分離之目的。此晶片以使用聚二甲基矽氧烷(polydimethylsiloxane, PDMS)為基材製造而成，由於PDMS本身疏水性不利於本研究之應用，故本研究用了多層次表面改質技術將親水並帶負電荷之聚合物材料附著於毛細管電泳晶片之管道上。常關式閥門於晶片上形成一條奈米管道(nanoscale channel)以連接樣品預濃縮區及分離管道，並於表面改質後產生帶負電荷之特性。此奈米管道只允許電流通過，因而帶負電荷之DNA便會被阻擋於閥門前，進而達成集中濃縮效果。預濃縮步驟完成後，施加一吸力氣壓將常關式閥門下壓，使預濃縮樣品區及分離管道接合。當電壓持續供應之下，已完成預濃縮之DNA便會快速的被驅動至分離管道，以進行後段的樣品分離與分析。此研究之晶片成功的達到DNA樣品預濃縮及分離效果。結果顯示樣品預濃縮區之螢光偵測訊號於1.9分鐘之預濃縮時間內有效的增強了大約3,570倍。與傳統之十字型毛細管電泳晶片比較，在預濃縮100秒的條件下，標準品X174 DNA/HaeIII之分離訊號成功的增強了41倍。這些結果顯示本晶片可成功的達到DNA樣品預濃縮及分離效果，進而成功的降低了毛細管電泳晶片之偵測極限。
英文摘要 A simple, sequential DNA pre-concentration and separation method by using a micro capillary electrophoresis (CE) chip integrated with a normally-closed valve which is activated by pneumatic suction has been developed in this study. The CE chip is fabricated using polydimethylsiloxane (PDMS). A surface treatment technique for coating a polymer bilayer with an anionic charge is applied to modify the surface of the microchannel. A normally-closed valve with anionic surface charges forms a nanoscale-like channel that only allows the passage of electric current but traps the DNA samples so that they are pre-concentrated. After the pre-concentration step, a pneumatic suction force is applied to the normally-closed valve. The channels are reconnected as the valve membrane is pressed down by the suction force. The DNA samples are then moved into a separation channel for further separation, detection and analysis. Successful DNA pre-concentration and separation has been achieved. Fluorescent intensity at the pre-concentration area is increased approximately 3,570 times within 1.9 minutes of operation. The signals from the separation of X174 DNA/HaeIII markers are enhanced approximately 41 times after 100 seconds of pre-concentration time, as compared with the results using a traditional cross-shape micro CE chip. These results clearly demonstrate that successful DNA pre-concentration for signal enhancement and separation analysis can be performed by using this new micro CE chip.
論文目次 Table of Contents

Abstract........................................I
摘要............................................III
誌謝............................................V
Table of Contents...............................VII
List of Figures.................................IX
Nomenclature....................................XIII

Chapter 1: Introduction
1-1 MEMS and Bio-MEMS technologies..............1
1-2 Capillary electrophoresis...................3
1-3 Micro capillary electrophoresis (CE) chip...4
1-4 Literature survey...........................5
1-4-1 Online sample pre-concentration...........5
1-5 Motivation and objectives...................8

Chapter 2: Theory and Design
2-1 Theory of micro CE chip.....................10
2-1-1 Electrophoretic mobility..................10
2-1-2 Electroosmotic flow.......................11
2-1-3 DNA electrophoresis.......................12
2-2 PDMS surface modification...................13
2-3 Design of the micro CE chip.................15

Chapter 3: Materials and Methods
3-1 Micro CE chip fabrication...................22
3-1-1 Photomask fabrication.....................22
3-1-2 Substrate cleaning........................22
3-1-3 Lithography process.......................23
3-1-4 PDMS casting..............................24
3-1-5 Chip bonding..............................26
3-2 Channel surface modification................27
3-3 Experimental Setup..........................28

Chapter 4: Results and Discussion
4-1 Surface modification........................34
4-2 DNA pre-concentration.......................35
4-3 DNA separation..............................38

Chapter 5: Conclusions and Future Work
5-1 Conclusions.................................48
5-2 Future work.................................49

References
Biography .......................................58
Publication List................................59
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