本研究以波導模態共振生物晶片作為基礎，將奈米金粒子修飾在生物晶片的波導層上，期望藉由波導模態共振效應與粒子電漿共振效應的結合能夠提升生物晶片在生物分子檢測上的表現。本研究首先利用有限元素模擬分析，研究波導模態共振效應與粒子電漿共振效應結合時的光學特性，再透過折射率實驗及生化分子檢測驗證模擬結果。
本研究建立了三維具有奈米金之波導模態共振式生物晶片模型，藉由改變入射角調整共振波段，在波導模態共振與粒子電漿效應間達到平衡，最終找到在共振波長540 nm時會具有最高的靈敏度。接著建立具有生物層結構之波導模態共振式生物晶片模型，以模擬三種不同檢測法之情形，分別為無奈米金傳統法、有奈米金傳統法、新型三明治法，模擬結果顯示以新型三明治法進行檢測時與有奈米金傳統法相比，光譜靈敏度、穿透光譜面積分別提升了79.33 %與157.24 %，而從漸逝波分佈與修飾結構來觀看，新型三明治法會使得能量會集中在奈米金與表面波導層之間，且由於其目標分析物之反應位置更靠近奈米金，因此更能夠有效利用粒子電漿共振效應。
實驗部分透過折射率實驗找到共振波長在540 nm時晶片會有最高的靈敏度，實驗結果與模擬分析結果相符；在生化分子檢測則是選擇免疫球蛋白G做為檢測分子，並以三明治結構進行有無奈米金檢測的比較，觀察粒子電漿共振對於生化分子檢測的影響，而以無奈米金進行檢測之檢測極限為7.78×10-7 g/ml；有奈米金則是4.56×10-8 g/ml，將檢測極限降低了一個級數，顯示粒子電漿共振效應能夠大幅提升波導模態共振式生物晶片之生物分子檢測能力。本研究透過模擬解釋了不同修飾法所帶來的影響、闡述其背後光學特性，在未來修飾法的選擇及改良上能夠有更明確的方向。

This research aims to modify gold nanoparticles (AuNPs) on the grating structures of guided mode resonance biosensors to enhance the surface sensitivity of guided-mode-resonance (GMR) optical biosensors. We expected that surface sensitivity of guided-mode-resonance (GMR) optical biosensors can be enhanced by particle-plasmon-resonance (PPR). This research uses finite element method (FEM) analysis to numerically study the coupling between the GMR and PPR, which are validated through refractive index experiments and bio-reaction experiments.
For the numerical simulations, we first build a three-dimensional numerical model for GMR biosensors with AuNPs. By changing incident angel to shift the GMR resonant wavelength, the optimal wavelength of 540 nm was found to achieve the highest sensitivity via properly coupling GMR and PPR effects. Then a three-dimensional model of GMR biosensors with biolayer was developed to study the sensing performance for three different models, namely the traditional method without AuNPs, traditional method with AuNPs, and new sandwich method. The simulation results show that sensitivity and intensity variety of new sandwich method are enhanced by factors of 79.33 % and 157.24 % compared with the traditional method with AuNPs. This enhancement can be explained by the evanescent wave distribution, which shows strong field enhancement occurs between AuNPs and waveguide surface without distorting the GMR, confirming enhanced bio-reaction PPR.
For experiments, in refractive index experiments, we find that the highest sensitivity occurs at a resonance wavelength 540 nm. In bio-experiment, IgG detection was conducted for sandwich structure to compare with without AuNPs and with AuNPs. The limit of detection (LoD) of without AuNPs and with AuNPs are 7.78×10-7 g/ml and 4.56×10-8 g/ml respectively, showing that PPR can enhance surface sensitivity of GMR optical biosensors. This study explains coupling between PPR and GMR and the optical properties, proving insights for the development of new GMR biosensors with AuNPs for sensitive bio-detection.

目錄
致謝 i
摘要 iii
Abstract iii
目錄 vi
圖目錄 ix
表目錄 xv
第一章 緒論 1
1-1 前言 1
1-2 生物感測器的種類 1
1-3 光學式生物感測器的發展 2
1-4 文獻回顧 7
1-4-1 波導模態共振式生物晶片 7
1-4-2 反射式波導模態共振生物感測器量測系統 11
1-4-3 以粒子電漿共振效應強化生物晶片檢測 14
1-4-4 三明治檢測法之文獻回顧 19
1-4-5 文獻總結 20
1-5 研究動機 21
1-6 論文架構 21
第二章 理論背景 23
2-1 波導理論 23
2-2 光柵繞射原理 24
2-3 漸逝波基本原理 26
2-4 波導模態共振理論 27
2-5 波導模態共振條件 28
2-6 粒子電漿共振(Particle Plasmon Resonance, PPR) 29
2-7 奈米金粒子特徵光譜特性 30
2-8 生物感測系統檢測原理 31
2-8-1 以共振波長飄移量作為檢測原理 31
2-8-2 以光強度變化作為檢測原理 32
第三章 生物晶片光學特性分析 35
3-1 前言 35
3-2 馬克斯威爾方程式 36
3-3 模型建立 36
3-3-1 幾何模型與網格建立 36
3-3-2 材料參數與邊界條件設定 39
3-4 奈米金粒子對共振波長之影響 41
3-4-1 奈米金分佈對光學特性之影響 46
3-5 建立生物層結構之GMR生物晶片模型 47
3-5-1 幾何模型與材料參數 48
3-6 奈米金位置與光譜之關係 49
3-7 奈米金位置與能量分佈之關係 52
3-8 奈米金位置與電場分布之關係 54
3-9 新型三明治法實際情況之計算 55
3-10有限元素模擬之結果與討論 57
第四章 具有奈米金之波導模態共振式生物晶片之製程與反射式光強度實驗 58
4-1 前言 58
4-2 波導模態共振式生物晶片之製作 58
4-2-1 波導層厚度的控制 59
4-3 奈米金粒子的製備與修飾 60
4-3-1 奈米金粒子合成步驟 61
4-3-2 修飾奈米金粒子於波導層之步驟 61
4-4 波導模態共振反射式光強度實驗 62
4-4-1 波導模態反射式共振光強度實驗流程 62
4-4-2 反射式光強度量測系統架構 63
4-4-3 反射式共振波長光強度實驗步驟 64
4-4-4 反射式共振波長光強度實驗結果 64
4-5 實驗結論與討論 67
第五章 反射式系統檢測生化分子 69
5-1 前言 69
5-2 免疫球蛋白G(Immunoglobulin G, IgG)簡介 70
5-3 生化分子檢測策略 70
5-4 溶液配置與實驗步驟 71
5-4-1 修飾Capture anti-IgG與IgG步驟 73
5-4-2 生物標記檢測步驟 74
5-5 多濃度檢測極限(Limit of detection, LoD)之計算 75
5-6 單濃度檢測極限之計算 77
5-7 免疫球蛋白G標準樣品檢測結果 77
第六章 結論與未來工作 80
6-1 結論 80
6-2 未來展望 81
6-2-1 以新型三明治法檢測IgG 81
6-2-2 改變光柵週期 81
6-2-3 以生物分子結構建立模型 82
第七章 參考文獻 84

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