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標題: | 同步壓印微米預濃縮流道與奈米狹縫表面電漿共振感測器用於免標定免疫分析 Synchronously Imprinting Preconcentrator and Nanoslit Surface Plasmon Resonance Sensor for Immunoassay |
作者: | Wen-Fei Yang 楊文斐 |
指導教授: | 沈弘俊(Horn-Jiunn Sheen) |
共同指導教授: | 魏培坤(Pei-Kuen Wei) |
關鍵字: | 深反應離子蝕刻,奈米壓印技術,電驅動奈米流體預濃縮,週期性奈米金屬表面電漿共振,生物感測器,免標定免疫分析,實驗室晶片, Deep Reactive Ion Etching,Nanoimprint Lithography,Electrokinetic-based Nanofluidic Preconcentration,Nanoslit-based Surface Plasmon Resonance,Lab on a chip, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 本研究目的為開發新型免疫分析檢測裝置,藉由奈米壓印將奈米流體預濃縮(Nanofluidic Preconcentration)微流道與週期性奈米金屬狹縫表面電漿共振(Surface Plasmon Resonance, SPR)感測器整合於同一片塑膠基板上。首先將待測活性分子進行預濃縮,再藉由調整電位差來控制濃縮區塊,使其準確移動於表面電漿共振晶片上方,最後量測其光譜訊號紅移(Redshift)以進行後續免疫分析。
矽晶圓母模製程,首先係利用電子束微影(E-beam Lithography)和反應離子蝕刻(Reactive Ion Etching, RIE)在矽晶圓基材上定義週期性奈米狹縫結構。接著使用正光阻作為作為乾蝕刻阻擋層,在奈米結構上方定義出微米流道遮罩,並使用感應耦合電漿離子蝕刻(Inductively Coupled Plasma Reactive Ion Etching, ICP-RIE)技術完成微米流道與奈米狹縫於同一片矽晶圓母模上。之後以環烯烴類聚合物(Cyclic Olefin Polymer, COP)作為晶片材料,與矽晶圓母模進行奈米熱壓印將結構轉印至此高分子聚合物上,經由遮罩濺鍍完成局部鍍金,再使用奈米多孔性材料Nafion®作為奈米流道鋪設於兩表面電漿共振晶片間,確立奈米預濃縮結構。最後使用聚二甲基矽氧烷(Polydimethylsiloxane, PDMS)作為覆蓋材料,經由化學表面修飾及氧電漿表面改質接合COP與PDMS,即完成本免標定免疫分析晶片。 本研究檢測之樣本為人類皰疹病毒第四型(Epstein-Barr virus, EBV)抗體,通入30 ng/mL的重組蛋白抗原(Latent Membrane Protein 1, LMP1)於濃縮流道內,藉由操控電壓使濃縮區塊限定於奈米狹縫晶片上,接著比對實驗組與對照組的穿透特性光譜,經由紅移量的差異和LMP1抗原濃度參考曲線,我們發現濃縮將30 ng/mL抗原濃度提升至約300 µg/mL,倍率達10,000倍,並藉此再將3 pg/mL濃度抗原進行濃縮,驗證得出其最小檢測濃度為3 pg/mL。 總結言之,此以奈米壓印技術成功達到量產與低成本的快速製程,且奈米流體預濃縮降低了檢測下限,加上表面電漿共振具有高靈敏度、即時性、和免標定的優勢,我們藉由簡便的量測系統完成了一個免標定且微量的超低濃度檢測平台。 In this study, an immunoassay platform that integrating nanofluidic preconcentrator with periodic metallic surface plasmon resonance sensor by nanoimprinting has been developed. The concentrated protein was trap in the sensing area of SPR in a microfluidic channel by electrical potential difference. Then, measuring the spectral signal red-shift of antibody-antigen interaction on periodic metallic slits for subsequent immunoassay. The periodic nano-grating structure were clarified and fabricated on a silicon wafer by E-beam lithography and reactive ion etching, then the microchannel mold were fabricated by inductively coupled plasma reactive ion etching. These structure was transferred onto a cyclic olefin polymer by nanoimprint lithography. The gold was deposited on the grating structure of COP by sputter. A porous material, Nafion, was used as the ion-selective channel, was aligned to the grating structure on COP. Overlay were made by PDMS. After the chemically modified surface treatment, the COP can be bond with PDMS by oxygen plasma. And the label-free immunoassay biochip has been done. Thereafter, Epstein-Barr Virus Antibody (EBV antibody) and Latent Membrane Protein 1 (LMP1) were used as the testing samples for future works. The result is that the concentration fold of LMP1 antigens can be raised up to approximately 10,000 folds as the 30 ng/mL of antigens was condensed to 300 µg/mL. And we successfully verify that the minimal detectable limit is about 3 pg/mL by concentrating 3 pg/mL of antigens to 30 ng/mL. In summary, by integrating preconcentrator and SPR system, we can have a label-free, preconcentration detectable platform by using simple measurement system. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77948 |
DOI: | 10.6342/NTU201702123 |
全文授權: | 有償授權 |
顯示於系所單位: | 應用力學研究所 |
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