表面電漿共振儀之動態相位偵測技術 與微量生物分子檢測應用

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姓名

謝佾儒(Yi-ru Hsieh) 查詢紙本館藏

畢業系所

光電科學與工程學系

論文名稱

表面電漿共振儀之動態相位偵測技術與微量生物分子檢測應用
(Dynamic Phase-Acquisition Technique and Low-Abundance Biomolecule Detection of a Surface-Plasmon-Resonance System)

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摘要(中)

本論文以研究團隊所研製之相位式表面電漿共振儀為基礎，開發動態相位偵測技術以提升系統折射率解析度後，並著手進行生物分子的實驗量測。目前，本系統之相位偵測穩定度在2000秒內可達0.5859度，所對應的系統折射率解析度為 RIU(Refraction Index Unit)。以本系統進行胎兒纖維結合素之抗體抗原專一性鍵結相位檢測，當抗原濃度為50 、2.5 、1.25 時，其相位變化分別為3.0154度、1.0613度、9.729度，並由四層結構系統進行數值擬合，擬合之結果得濃度為50 時，鍵結等效折射率為1.33318，等效厚度為10.81 nm；濃度為2.5 時，擬合之等效折射率為1.33297，等效厚度為10.43 nm；濃度為1.25 時，擬合之等效折射率為1.33227，等效厚度為9.84 nm。可知相位訊號、等效折射率及等效厚度均隨抗原濃度變化，有機會經由系統校正程序獲得關係曲線後，應用此系統進行抗原濃度檢測。

摘要(英)

In this thesis, a dynamic phase-detection technique was developed to improve the refractive-index resolution of a surface-plasmon-resonance system previously built in the research group. After examining the system stability, biological molecule measurements were performed. The phase-detection stability was 0.5859 degree in 2000-second period, and the corresponding refractive-index resolution was 1.3×10-5 RIU (Refraction Index Unit). This system was applied to antibody-antigen binding experiments of fetal fibronectin. With antigen concentrations of 50 ng/ml, 2.5 ng/ml, and 1.25 ng/ml, the phase changes were 3.0154, 1.0613 and 9.729 degrees, respectively. Considering a four-layer Kretschmann configuration, the equivalent thickness and the equivalent refractive index of the antibody-antigen binding layer were found by numerical fitting. For the concentration of 50 ng/ml, the equivalent refractive index was 1.33318 RIU and the equivalent thickness was 10.81 nm. For the concentration of 2.5 ng/ml, the equivalent refractive index was 1.33297 RIU and the equivalent thickness was 10.43 nm. For the concentration of 1.25 ng/ml, the equivalent refractive index was 1.33227 RIU and the equivalent thickness was 9.84 nm. As the phase signal, the equivalent refractive index and the equivalent thickness vary with the antigen concentrations, it is possible to obtain their relation curves through calibrations and then apply the system to determinations of the antigen concentration.

關鍵字(中)

★ 表面電漿共振系統
★ 胎兒纖維結合素

關鍵字(英)

★ Fetal fibronectin
★ Surface-Plasmon-Resonance system

論文目次

摘要 I
ABSTRACT II
誌謝 III
目錄 V
圖目錄 VIII
表目錄 XIV
第一章緒論 1
1.1 前言 1
1.2 文獻回顧 2
1.2.1 生物感測器-表面電漿共振儀 2
1.2.2 SPR 免疫檢測技術 8
1.2.3 胚胎纖維蛋白質 10
1.3 研究動機 11
1.4 論文架構 11
第二章表面電漿原理 13
2.1 表面電漿之色散關係式 14
2.2 SPR 產生方式 18
2.3 Kretschmann 組態-SPR 反射率及相位演算 21
2.3.1 兩層介質系統之反射係數 21
2.3.2 三層介質系統之反射係數 24
2.3.3 四層結構系統 27
2.4 小結 28
第三章光學解相理論與系統架構 29
3.1 光學干涉術原理 29
3.2 相移干涉術 32
第四章相位式表面電漿共振儀 36
4.1 相位式表面電漿共振儀 36
4.2 表面電漿共振儀架構 43
4.3 PZT回饋控制 48
4.4 感測片設計 52
4.5 程式設計 60
第五章實驗方法與實驗結果分析 67
5.1 實驗藥品 67
5.2 實驗方法 68
5.2.1 PBS 緩衝液配製 68
5.2.2 蛋白質配製 68
5.2.3 活性劑配製 68
5.2.4 牛血清蛋白配製 69
5.2.5 SPR 裝置 69
5.2.6 胎兒纖維蛋白質抗體固定化 70
5.3 實驗結果討論與模擬分析 72
5.3.1 實驗一 P極化光的量測 72
5.3.2 實驗二 Open-Mode系統穩定度量測 75
5.3.3 實驗三 Close Mode 系統穩定度量測 93
5.3.4 實驗四純水交換PBS系統相位變化量測 94
5.3.5 實驗五胎兒纖維結合素抗體抗原專一性鍵結實驗 97
第六章結論 103
參考文獻 104

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指導教授

李正中、陳怡君
(Cheng-Chung Lee、Yi-Chun Chen)

審核日期

2010-10-19

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