臺灣博碩士論文加值系統

本研究旨在發展一種新型的蛋白質微陣列(protein microarray)方法，包含使用微機電技術製作壓印晶片，並將蛋白質平行蓋印至生物檢測晶片上以形成蛋白質微陣列。本研究完成兩代壓印晶片之研發，第一代的壓印晶片為沾印式。沾印式壓印晶片利用微印章陣列以及微印台所構成之微印章-微印台壓印系統，可將不同之蛋白質平行轉印至生物檢測晶片之表面上。本研究之微印章係以彈性材料(polydimethylsiloxane：PDMS)製作而得，因此，微印章具有足夠之柔軟性可與粗糙平面做良好完整之接觸。實驗證明，本研究之沾印式壓印晶片可成功地將蛋白質溶液轉印至可結合蛋白質之表面，壓印出的蛋白質印之間的尺寸差異小於10%。但因表面張力作用關係，所能打印之尺寸難以小於350μm，因此無法達到高密度微陣列之需求。
為解決以上問題，本研究研發的第二代壓印晶片為背後填充式，填充式壓印晶片包含三個部分：上層為儲液區、中層是垂直微通道陣列區、下層是微印章陣列區。單一微印章之中心包含一微通道，生醫溶液注入儲液區，藉由表面張力驅動而流經中層微通道及微印章的微通道，最後來到微印章之壓印面，再與生物檢測晶片接觸，將生醫溶液傳送到生物檢測晶片上。為了製作出能產生功能之微印章，本研究同時進行數值模擬計算以求最佳的設計成果。由於驅動液體流動的總表面張力取決於液體-氣體界面之形狀，因此，數值模式必須具備能夠精確地追蹤液體-氣體界面的功能。本研究之數值模式採用流體體積法(Volume of Fluid, VOF)。第二代填充式壓印晶片的壓印實驗證明，蛋白質溶液可成功地轉印至生物檢測晶片上，且壓印出的蛋白質印亮度及面積之標準差可小於3%。此外，所能成功轉印之蛋白質印的尺寸可小至100μm。

摘要 I
致謝辭 iii
目錄 iv
圖目 vii
表目 xii
第1章 緒論 1
1-1 研究背景 1
1-2 微陣列技術 2
1-2-1 點針法 3
1-2-2 光學微影法 5
1-2-3 微噴墨頭噴射法 7
1-2-4 電噴灑法 9
1-2-5 微壓印法 11
1-2-6 凝膠方陣法 14
1-2-7 微流體通道法 18
1-2-8 蘸水筆奈米微影法 20
1-3 研究目的 22
1-4 本實驗室之壓印系統的設計及操作原理 23
第2章 文獻回顧 26
第3章 第一代沾印式壓印晶片之實驗製程與材料 30
3-1 微印章-印台系統之設計及操作原理 31
3-2 沾印式壓印晶片之製程 34
第4章 第一代沾印式壓印晶片之實驗結果與討論 41
4-1 PVDF膜微壓印實驗 41
4-2 PhastGel®上的微壓印實驗 45
4-3 APTS-BS3生物檢測晶片上的微壓印實驗 48
4-3-1 微壓印實驗設計 48
4-3-2 壓印尺寸之一致性 49
4-3-3 蛋白質與生物檢測晶片之間的結合強度 49
第5章 第二代填充式壓印晶片之實驗製程與材料 55
5-1 填充式壓印晶片之設計及操作原理 55
5-2 填充式壓印晶片之製程 60
5-2-1 填充式壓印晶片之第一代製程 60
5-2-2 填充式壓印晶片之第二代製程 75
第6章 第二代填充式壓印晶片之壓印實驗結果與討論 85
第7章 第二代填充式壓印晶片之數值模擬 92
7-1 數值模擬原理 92
7-2 蛋白質物理性質測量 97
7-3 工作流體為低濃度之蛋白質溶液或水的模擬結果與討論 99
7-3-1 格點系統及邊界條件設定 99
7-3-2 出水現象 101
7-3-3 靜態壓印尺寸 109
7-3-4 動態壓印過程 112
7-4 工作流體為高濃度之蛋白質溶液的模擬結果與討論 123
7-4-1 格點系統及邊界條件設定 123
7-4-2 蛋白質溶液壓印過程 127
7-4-3 影響壓印尺寸之因素 133
第8章 結論 149
參考文獻 151
附錄A 微印台陣列之製程 156
附錄B 生物檢測晶片之製程 159
附錄C ELISA-like之製程 162
附錄D 蛋白質微陣列之影像處理程序 164
附錄E SU8光阻之微影製程參數 165
附錄E-1 SU8-5，60μm之製程參數 165
附錄E-2 SU8-50，110μm之製程參數 166
附錄E-3 SU8-2005，20μm之製程參數 167
附錄E-4 SU8-2050，110μm之製程參數 168

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