臺灣博碩士論文加值系統

本研究應用機構運動方式完成新式的PCR反應系統，主要為設計與製造生物晶片之微加熱器控制與循環系統，目標是希望能將此系統運用在聚合酶連鎖反應(polymerase chain reaction, PCR)系統中以提供穩定之熱源。主要研究項目為：1.在載玻片上製作微加熱器並研究其效能與穩定度、2.微加熱器電源供給之電路設計與控制、3.循環機構之研製、4.循環機構之控制、5.反應槽製作。
利用微機電系統(Micro-Electro-Mechanical-System)技術在載玻片上製作出微加熱器，再使用本研究所製作之電壓調節電源供給系統以控制微加熱器溫度，並藉K-Type (TP-01Z)熱電隅、LabVIEW7.1軟體、及NI USB-9211資料擷取設備以測量與記錄微加熱器溫度隨時間變化之情形。結果顯示微加熱器之穩定性良好，最大溫度差異小於1°C。接著應用旋轉機構以達成PCR熱循環系統，並使用LabVIEW7.1軟體、及NI USB-6009 (Digital Output)訊號擷取卡，配合機構控制電路做訊號轉換，控制機構旋轉的時間。
本研究以三個微加熱器建立PCR所需的三個溫度，再以機構循環地移動檢體使其與不同溫度之微加熱器接觸，藉著熱傳導方式傳遞熱量而使檢體達到所需的反應溫度，成功應用機構式熱循環系統，製作出一新式的PCR反應系統。
機構式熱循環系統之反應槽內檢體的溫度上升速率約1°C/sec，下降速率為2.8 °C/sec，上升速率與傳統固定式PCR相比較為慢，原因是微加熱器與反應槽之間以接觸式熱傳導傳遞熱量，所以存在一接觸熱阻，此問題值得進一步探討。

The design and fabrication of a thermal system with voltage regulated micro heaters are presented. The thermal system is intended to provide stable heat sources to the polymerase chain reaction (PCR) system. This study is concentrated on (i) the design, fabrication and test of micro heaters, (ii) the circuit design and control of power supply for heaters, (iii) the fabrication and control of a rotary mechanism, and (iv) the fabrication of a reaction well.
By using the micro-electro-mechanical-system (MEMS) technology, micro heaters are fabricated on a microscope slide. The temperature of heaters is then adjusted by a voltage regulator. The variation of heater temperature with time is measured and recorded by a homemade data acquisition system consisting of K-type thermocouple, LabVIEW7.1 software, and NI USB-9211 data acquisition card. Results show that the stability of micro heaters is very good and the maximum temperature deviation is within 1°C. The thermal cycling system is further implemented with a rotary mechanism. The signal and operation of the mechanism are controlled by LabVIEW7.1 software and NI USB-6009 data acquisition card.
Three micro heaters are used to build up three temperatures that PCR needs. The sample is then rotated by the mechanism to contact with the micro heaters of different temperatures so that it achieves the required temperatures via heat conduction. A new PCR system is successfully developed in this study.
The sample temperature in the reaction well of the thermal cycling system rises at a rate of 1°C/sec and drops at 2.8 °C/sec. The rise rate is slower than that of the traditional PCR because there is a thermal resistance between the micro-heaters and reaction well as heat transferred by conduction. The problem is worth studying further.

摘 要 IV
ABSTRACT V
誌 謝 VII
目 次 VIII
表目錄 XI
圖目錄 XII
第一章 緒 論 1
1.1 前言 1
1.2 文獻回顧 2
1.3 研究動機與目的 5
第二章 聚合酶連鎖反應與接觸式加熱 6
2.1 DNA的組成與結構 6
2.2 聚合酶連鎖反應 7
2.2.1 PCR歷史與發展 7
2.2.2 PCR反應需具備的要件和材料 8
2.2.3 PCR的反應過程 9
2.3 PCR儀器設計方案 10
2.4 接觸式加熱基礎理論 14
第三章 儀器與溫控元件製作 16
3.1 儀器總架構 16
3.2 電源供給電路 16
3.3 溫度量測程式 19
3.4 LABVIEW軟體與硬體架設 20
第四章 微加熱晶片的設計與製作 22
4.1 晶片設計 22
4.2 光罩設計 24
4.3 晶片製程 25
4.3.1 晶片前處理 25
4.3.2 圖形定義 27
4.3.3 蒸鍍金屬 30
4.3.4 掀去製程 31
4.4 晶片接線與安裝 32
4.4.1 晶片接線 32
4.4.2 晶片固定安裝 33
第五章 反應槽設計與製作 36
5.1 反應槽需求與測試 36
5.1.1 實驗設備與方法 36
5.1.2反應槽幾何尺寸 38
5.1.3反應槽材料 38
5.2反應槽之設計製作 39
第六章 機構設計與製作 42
6.1 機構需求與構想設計 42
6.2 方案裁決與機構製作 43
6.2.1 整體設計 43
6.2.2 機構運動 44
6.2.3 動力設計 47
6.2.4 夾具設計 49
6.3 微加熱晶片支承結構 51
6.4 機構控制元件與測試 52
6.4.1 元件製作與整合 52
6.4.2 機構控制程式 53
6.4.3 運動測試 54
第七章 結果與討論 56
7.1 溫度設定前的討論 56
7.1.1微加熱器熱應力破壞與改善 56
7.1.2微加熱器與反應槽之間隙修正 58
7.2 微加熱器溫度設定方法與步驟 59
7.3 微加熱器實際量測 60
7.4 PCR溫度循環 66
7.4.1 連續溫度變化結果 66
7.4.2 溫度循環中降溫之改進 66
7.4.3 改良後之溫度循環結果 67
7.5 機構式熱循環PCR系統實測 70
第八章 結論與未來展望 71
8.1 結論 71
8.2 未來工作 72
參考文獻 73
作者簡介 76

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