在現今的生醫晶片系統中，量測人體全血中的尿素和血糖是健康相當重要的指標。血尿素氮 ( BUN) 的形成主要是來自蛋白質代謝的最終產物，因此血中尿素氮的濃度，也可以用來做評估腎功能的指標，所以當血液中的尿素氮值升高，超過標準值時表示腎臟過濾功能可能出現問題，這是一個重要的警訊。然而，隨著現今全球文明化和飲食文化的改變，許多的文明病便相伴而至例如糖尿病，所以血糖指數的多寡是相當重要的，當血糖值過高表示胰島素的缺乏是糖尿病的高危險群，因此開發一個生醫晶片整和電化學電泳 ( CE-EC )去量測生醫樣品，以期能運用在醫療檢驗上做為疾病的預防和檢測機制是相當重要的。
本研究提出了一種新的方法來檢測人體全血中的尿素和葡萄糖，利用多種不同反應的酶滴在線微流體上使其和檢測樣品做反應以利偵測，再者使用PVC膜包覆在線體的外部形成一個密閉式管道。滴在線上的酶可以直接和樣品反應無須經過任何的表面改質和加工，而流經過的生物樣品經由酶的轉化，最後再由下端的電極做電化學檢測。所以利用以上的方法結合CE -EC檢測生物樣品，可以得到很好的量測結果。本研究進一步做人體的全血量測，首先利用RBC裂解液滴在多出的注入端部分而後在加入約2-µL全血混和後，利用正負高電壓驅動線微流體上的電滲流流動，可以裂解血液中的紅細胞和同時過濾掉其他雜質，做為一個過濾器使用，使其提高偵測性能和對阻塞管道的影響降低。由於焦耳熱效應關係，管道上緩衝液和樣品是很容易蒸發，而且溫度上也不耐高，容易造成線體的損壞，所以塗佈PVC 膜可以改善以上的情形。此外，PVC塗佈的線體可以在更高的分離電場500 V/cm進行操作。
一般尿素和飯前血糖的正常範圍值分別是1.78~7.12 Mm 和 3.89~6.11 mM ，結果表示，此研發晶片系統在量測表準樣品尿素和葡萄糖中擁有良好的線性結果，尿素濃度範圍在0.1 mM – 10.0 mM (R2=0.9850) 間；葡萄糖濃度範圍在0.1 mM – 13.0 mM (R2=0.9668) 間，擁有良好的靈敏度。在全血的檢測中也成功分離出血液中的離子，並測得尿素和血糖濃度分別為3.98 mM 和 4.94 mM。本研究開發的線微流體系統提供了一個低成本，高性能且創新的方法應用在人體血液中的檢驗。

In biological chip systems, urea and glucose are important indicators for detection in human blood. Urea forms in the liver as a waste product of protein metabolism then collects in the bloodstream. The concentration of blood urea nitrogen (BUN) is an important biological indicator used for monitoring the functionality of the kidney. In general, patients with kidney failure have high BUN levels. However, with changing food culture, glucose has also become a identify control factor for diabetes mellitus. Excessive glucose displays a deficiency of insulin. Therefore, developing a microfluidic device for capillary electrophoresis electrochemical (CE-EC) detection of bio-samples is a necessary.
This research presents a novel technique for detection in human whole blood which utilizes a novel enzyme-doped thread with a PVC (polyvinylchloride) membrane coating for on-site urea and glucose detection on a thread-based microfluidic device. The enzyme can be directly applied to the thread without delicate pretreatment or a surface modification process. The passing biomolecules are digested by the enzymes and then electrochemically detected downstream. With this approach, CE-EC detection with on-site bio-reaction can be simply achieved. The whole blood sample is first mixed with a RBC lysis buffer to prevent blood coagulation. The lysed RBC and other solid pieces are simultaneously filtered away while electrokinetically flowing through the thread-based microfluidic system. A thin layer of PVC membrane is coated on the enzyme-doped thread to further fix the applied enzyme and to prevent rapid evaporation of the running buffer due to the Joule heating effect. In addition, the PVC coated thread can be operated at a higher separation electric field of 500 V/cm due to the reduction of buffer evaporation.
Results also indicate that the developed system exhibits a good linear dynamic range for detecting urea and glucose in concentrations from 0.1 mM – 10.0 mM (R2=0.9850) and 0.1 mM – 13.0 mM (R2=0.9668), which is suitable for adoption in detecting the BUN concentration in serum (1.78~7.12 mM) and the glucose fasting measuring range (3.89~6.11 mM). The whole blood detection shows that the developed thread-based microfluidic system can successfully separate the ions, BUN and glucose in blood. The calculated concentrations for BUN and GLU-AC in the whole blood sample are 3.98 mM and 4.94 mM, respectively.

Acknowledgements I
中文摘要 III
Abstract V
Table of Contents VII
List of Figures X
List of Tables XIII
Nomenclature XIV
Abbreviations XVI
Chapter 1 Inotoduction 1
1.1 Background 1
1.2 Capillary electrophoresis 1
1.2.1 The basic principle of electrophoresis 1
1.2.2 Substrate and production methods of electrophoresis chip 5
1.2.3 Detection methods of electrophoresis chip 7
1.3 Electrochemical 9
1.3.1 The system of electrochemical electrode 9
1.3.2 Detection methods of electrochemical 10
1.4 Paper microfluidic chip 13
1.5 Motivation and objective 16
1.6 Thesisorganization 17
Chapter 2 Theory and Design 19
2.1 Characteristics of thread 19
2.1.1 Hydrophilic and capillary force 19
2.1.2 Characteristics of polyester thread 20
2.2 Explore 2D and 3D electrodes 21
2.3 Biomicrofluidic chip 22
Chapter 3 Methods and Materials 25
3.1 Chips design 25
3.1.1 3D detecting electrode chip 25
3.1.2 Enzyme-doped thread coated with PVC membrane chip 26
3.2 Biochips fabrication 28
3.2.1 3D electrodes fabrication 28
3.2.2 Enzyme-doped thread coated with PVC membrane fabrication 29
3.3 Explore the surface of the plasma-treated 31
3.4 Explore PVC membrane coated with thread 32
3.5 Variable sample volumes 33
3.6 Reagent and apparatus preparation 34
3.6.1 Regent preparation 34
3.6.2 Experimental apparatus 35
3.7 Experimental procedure 36
3.7.1 Fundamental measurements 36
3.7.2 Biosample detection 37
3.7.3 Human whole blood detection 38
Chapter 4 Results and Discussion 40
4.1 EOF characteristics of the thread-based microfluidic system 40
4.2 Explore the variable volumes on thread 41
4.3 Evaluate concentration of enzyme 45
4.4 Performance evaluation on the2D and 3D electrodes 47
4.5 Electrophoresis of enzyme doped thread coated with PVC membrane 49
4.6 Detection of human whole blood on thread microfluidic system 50
4.7 Detection of human whole blood with enzyme doped thread coated with PVC membrane 52
Chapter 5 Conclusions and Future Work 55
5.1 Conclusions 55
5.2 Future work 56
Reference 57
Biography 66
Publication list 66

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