發展一個可靠、有效並且拋棄式的感測裝置，對於無論是在醫師早期診斷疾病或是一般大眾在家檢測疾病相關的因子都是極有幫助的。重要的是感測裝置的價錢決定了民眾是否可以負擔得起這種醫療照護，而操作的簡易度決定了這個裝置是否可以被應用在資源貧乏的地區。在過去發展平價裝置的研究中，有一部份致力於利用高分子材料(如：聚二甲基矽氧烷)製作平價裝置，值得一提的是紙基分析裝置在發展平價裝置的研究中展現出亮眼的成果。然而，在過去紙基分析裝置的研究中對於核酸檢測的相關研究仍然相對匱乏，導致紙基分析裝置應用在診斷病毒相關疾病時受到限制。在此，我們想要發展一個簡單且有效的方法在紙基分析裝置上檢測特定的核酸序列並且得到定量分析的結果，藉由完成以下步驟： 1) 我們利用一種叫做反轉錄環介導等溫擴增的分子生物技術，在同一溫度下增幅病毒的特定核酸序列以提高核酸濃度用於檢測，並且在紙基平台上利用螢光染劑做為檢測結果的呈色。這部分的研究可幫助我們發展一個可用於檢測病毒相關疾病的紙基檢驗法。 2) 我們發展了一個可以與智慧型手機整合的拋棄式的螢光取像裝置，用於分析先前的病毒檢測實驗中所得到的螢光影像。我們試著去證實裝置在截取不同螢光波長影像的準確性，且在不影響這個拋棄式螢光取像裝置的性能下，簡化裝置所需的元件，藉此達到降低成本且使紙基分析裝置也能利用螢光呈色反應。在所有的研究架構下，我們希望這些研究成果在未來可以達到利用智慧型手機以及紙基裝置做到準確檢測病毒相關的疾病。而在本研究論文的最後，我們也探討紙基分析平台應用於自動化液體操作機器的可行性，希望未來可以將紙基平台用於大量自動化篩選化學分子的實驗中，利用簡單便宜的材料，減少化學分子篩選過程中所需的成本花費。

The development of a reliable, effective and portable sensor is useful no matter for doctors to diagnose disease at early stage, or for the general public to monitor the amount of disease-relating factor at home. Most importantly, the price of sensor decides whether people can afford for this healthcare, and also the device operation decides whether this product can be used in resource limited area. Previous studies of developing low-cost devices partially focused on the polymer (i.e., PDMS) made device, and it is worth noting that the paper-based analytical device (PAD) showed outstanding achievement of the low-cost device development. However, the studies of paper-based analytical device were lack of nucleotides based assay that restricted the application of diagnosing virus related diseases. Here, we would like to develop a simple and efficient approach for detecting specific sequence of nucleic acids on paper-based analytical device with analyzed reports via the following procedures: 1) We employed a molecular biotechnology called RT-LAMP (reverse transcription loop-mediated isothermal amplification) for amplifying the specific sequence of nucleic acids from viruses at a constant temperature (dengue virus serotype-2 RNA in this study) and to utilize the fluorescent probes for revealing the detecting results on paper-based platform. This part helps us to develop a paper-based assay that could detect virus relevant disease. 2) We developed a portable fluorescent image recording device that is compatible with smart phone, in order to analyze the fluorescent images that are captured from the results of virus detecting assays. We tried to validate the accuracy in different fluorescent wave length, and simplified the components without affecting the performance in this portable fluorescence recording device. To take together, we hope these studies can achieve a goal that using a smart phone and a paper-based device to diagnose the virus-derived diseases. At the end of this dissertation, we also investigated the feasibility of applying paper-based platform on liquid-handling robot for the screen of specific chemical compound. Hope in the future, a paper-based platform can be employed on massive screening of chemical compounds in order to diminish the cost of chemical selection.

致 謝 3
中 文 摘 要 4
Abstract 5
Chapter 1
Introduction 7
1.1 Paper selection 9
1.2 Fabrication technique 11
1.2.1 Photolithography 12
1.2.2 Flexography printing 12
1.2.3 Wax related fabrication 13
1.2.4 Inkjet etching and plasma treatment 14
1.2.5 Screen printing and inkjet printing 15
1.2.6 Cutting technique 15
1.2.7 Three-dimensional (3D) paper-based microfluidic devices 16
1.3 Quantitative analysis 19
1.3.1 Colorimetric detection 19
1.3.2 Electrochemical detection 22
1.3.3 Luminescent detection 23
1.3.4 Fluorescence detection 25
1.3.5 Electrical conductivity measurement 26
1.4 Application areas 27
1.4.1 Environmental monitoring 27
1.4.2 Food safety 28
1.4.3 Health diagnostics 29
1.5 Overview of this dissertation 31
Chapter 2
Molecular-level dengue fever diagnostic devices made out of paper 35
2.1 Introduction 35
2.2 Materials and methods 38
2.3 Results and discussion 39
2.3.1 Strategies for serotype-2 dengue fever diagnosis in paper 39
2.3.2 Screening the appropriate fluorescence probe used in paper 43
2.3.3 Fluorescence-based examinations of the RT-LAMP products 46
2.4 Conclusions 50
2.5 Chapter 2 Appendix 51
Chapter 3
Portable fluorescence image recording device combined with paper-based platform 53
3.1 Introduction 53
3.2 Materials and Methods 54
3.2.1 Chemicals and materials 54
3.2.2 Portable fluorescent image recording device 55
3.2.3 Image capture and intensity analysis 55
3.3 Results and discussion 56
3.3.1 Design of portable image-recording device 56
3.3.2 Fluorescence detection via portable image recording device 59
3.4 Conclusions 62
Chapter 4
An approach to enhance self-compensation capability in paper-based devices
for chemical sensing 63
4.1 Introduction 63
4.2 Materials and methods 66
4.2.1 Chemicals and materials 66
4.2.2 Tolerance analysing 66
4.2.3 Colorimetric assay 67
4.2.4 Data analysis 68
4.3 Results and discussion 68
4.3.1 Performance of liquid-handling robot 68
4.3.2 Strategy for compensating droplet location 72
4.4 Conclusions 81
Chapter 5
Future works 83
5.1 Amplifying concentrations of DNA in serum or blood by RT-LAMP 83
5.2 Raising the reliability of portable fluorescent image recording device 83
5.3 Increasing available types of fluorescent probe on paper-based device 85
Reference 87
List of publications 100

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