Application of Surface Plasmon Resonance sensing for protein immunoassay in a micro/nano-fluidic chip

表面電漿共振技術近年來被大量應用在發展生物分子感測器上，這是一種利用光學的方式，便可達到非標記、高敏感度、少量樣本、即時的檢驗方法，利用生物分子免疫分析的特殊選擇性，可以在複雜的混合物中偵測到相當低濃度的特定分子。本研究著重於光學系統架設與應用，先從理論與模擬開始探討，利用稜鏡耦合的方式激發表面電漿波，進而建立一組可靠的生物分子感測系統，使用不同濃度的蔗糖水溶液對系統進行測試其靈敏度147.34◦/RIU與解析度(每個像素：3.5×10-5RIU)，最後應用於微奈米流道晶片，利用微奈米流道尺寸上的特性，提高分子偵測的濃度範圍，從蛋白質免疫分析實驗可知SPR角度的偏移在普通金膜實驗上為0.1°而在微流道晶片中增加至1.1°。Surface plasmon resonance (SPR) is a kind of optical detecting method and popular using in biosensor recent years. Compared with other technologies, SPR biosensor has several advantages such as real-time monitoring, label-free detection and small sample volume .Compare with the immunoassay of biomolecule , SPR sensing could provide high specificity and sensitivity in detect analytes in complex biological media. In this paper, we focus on SPR optical system’s building and application of biosensing. We discuss SPR from theory to simulation and build a SPR biosensing system by using prism coupler mode to excite Surface plasmon wave. Then we use different concentration of sucrose solution to test our SPR system and provide a good system sensitivity(147.34◦/RIU) and resolution(each CCD pixel : 3.5×10-5RIU). At last, we combine SPR system with micro/nano-fluidic chip and use the advantage of the micro channel size to enhance the detection range of biomolecule immunoassay. From the result of protein immunoassay experiment, we find out the SPR shift angle is 0.1° in bulk gold chip and it raise increase to 1.1° in micro/nano- fluidic chip.致謝.............................................II文摘要....................................IIIBSTRACT.....................................IV錄........................................V目錄....................................VII目錄......................................XI一章 導論...............................11.1 研究背景.............................11.2 研究動機與目的......................41.3 本文結構............................5二章 實驗理論與模擬....................62.1表面電漿理論.........................62.2 相關文獻回顧........................122.3稜鏡耦合組態之原理...................162.4 SPR角度計算與模擬...................19三章 實驗材料與方法....................233.1 實驗系統架設........................243.2金屬膜與稜鏡的選擇與製備.............27 3.2.1 稜鏡的選擇.......................27 3.2.2 金屬膜的選擇與製備...............293.3 待測樣本的製備......................34 3.3.1 蔗糖水溶液及折射率分析...........34 3.3.2 待測蛋白質的分子固化流程.........353.4 SPR訊號量測方式與分析方法...........403.5 微奈米流道晶片......................43四章 實驗結果與討論....................454.1普通金膜晶片之訊號分析結果............45 4.1.1 蔗糖水溶液之訊號分析..............45 4.1.2 蛋白質樣本之訊號分析..............494.2 微流道晶片之訊號分析結果.............54 4.2.1 蔗糖水溶液之訊號分析...............55 4.2.2 蛋白質樣本之訊號分析..............58五章 結論與未來展望.....................59考文獻...................................6