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研究生:陳怡君
研究生(外文):Yi-Chun Chen
論文名稱:利用光學消逝波偵測的新型生物晶片掃瞄系統
論文名稱(外文):A Novel Microarray Scanner Based on Evanescent Wave Excitation
指導教授:莊曜宇
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:電機工程學研究所
學門:工程學門
學類:電資工程學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:64
中文關鍵詞:生物晶片光學消逝波
外文關鍵詞:MicroarrayEvanescent Wave
相關次數:
  • 被引用被引用:1
  • 點閱點閱:266
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:2
在偵測基因微陣列晶片的系統中,如何能快速、大面積偵測,並同時避免背景光的影響是發展微陣列晶片偵測最重要的課題,在此研究中我們建構一新的光學激發及讀取系統,可以大面積並且靈敏的檢測在玻璃玻片上微陣列之螢光訊號。
我們使用白光光源為激發光源,因為白光光源為一廣波段的光源,配合不同波段的濾鏡就可以使用不同波段的光源,不但方便且濾鏡價格也比更換雷射光源低得多。以線型光纖導光器將白光光源導成線型輸出,利用光纖耦合法將此線型光源導入玻璃片內,利用玻璃與空氣介面之全反射作用在玻璃表面上產生均勻大面積的平面光學消逝波,以此平面光學消逝波作為激發源,激發在玻璃片上的DNA微陣列,經過透鏡、濾光片,以CCD讀取此微陣列螢光訊號。由於此系統利用平面光學消逝波為激發源,可避免一般以CCD作為偵測系統時,因激發光造成的背景雜訊。我們用程式控制移動平台、CCD及照片接合,達成自動化且快速的讀取整個DNA微陣列晶片。
我們比較使用此系統與市售儀器所得之DNA微陣列晶片影像,影像皆相符且可清楚分辨螢光強度差異。一般在玻璃等透明基材上的DNA微陣列可由此系統快速、靈敏的讀出,我們並將此系統應用於非螢光的偵測,例如以奈米金粒子取代螢光物質做為訊號來源的晶片或是細胞晶片等。
In this thesis, we present a high throughtput microarray reading system for detecting fluorescent signals on a glass slide. Moreover, cost of our microarray reader is much lower than that of commercial instruments due to its simple design.
Different from conventional laser scanning microarray readers, we used a white light (metal halide) as the excitation light source. It provided large-area illumination and flexibility of changing excitation wavelengths by using various optical filters. The white light was coupled into a glass chip by using a fiber optic dual lines light guide. As the light propagated in the glass waveguide, total internal reflection at the glass-air interface produced evanescent planar waves (EPW). The EPW excited fluorescence dyes in the close proximity of the chip surface. The fluorescence signals were then taken by an objective lens and a charge coupled device (CCD). We developed a Visual Basic program to take a set of fluorescent images at different positions. The whole image of the DNA microarray was obtained by image combination.
The reader has a spatial resolution of about 3.5um. The test of Cy3 fluorescence showed concentration below 10pM can be imaged within 10 seconds. However, the background of this system was larger than that of the confocal laser scanning method. It may be due to fluorescence of the glass and scattering of surface defects. Because of the novel EPW excitation design, our system is also useful for DNA microarrays labeled with gold nanoparticles and label-free cell arrays.
摘要 I
Abstract II
致謝 III
目錄 IV
圖目錄 V
表目錄 VII
第一章 微陣列晶片(Microarray) 1
1.1. 簡介 1
1.2.微陣列晶片的種類 3
1.2.1. 基因微陣列晶片 (DNA Microarray) 3
1.2.2. 蛋白質微陣列晶片(Protein Microarray) 4
1.2.3. 細胞陣列晶片(Cell Array) 5
第二章 微陣列晶片的偵測技術 6
2.1. 基因晶片的偵測儀器 6
2.1.1. 雷射掃瞄、PMT偵測 7
2.1.2. 均勻白光激發、CCD偵測法 9
2.2. 研究目的與方向 10
第三章 本研究所用之激發源-光學消逝波 11
3.1. 簡介基因微陣列晶片的螢光激發技術 11
3.2. Evanescent Planar Wave Technology 17
3.3. 光纖耦合法(Fiber coupling) 19
第四章 實驗架構 23
4.1. 實驗材料 23
4.2. 偵測系統之架構 25
4.3. 系統之解析度 28
第五章 結果與討論 33
5.1. 耦合效率(Coupling Efficiency) 33
5.2. 玻片周圍散射光的影響 34
5.3. 電動平移台之修正 36
5.4. 訊號分析 38
5.5. 背景值之探討 41
5.6. 結論及應用 43
參考資料 47
附錄1. Visual Basic 系統控制程式 49
附錄2. Matlab 圖像轉檔程式 64
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