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標題: | 利用自動化奈米壓印技術製作奈米狹縫表面電漿共振晶片用於鼻咽癌的DNA檢測之研究 Nanoslit Surface Plasmon Resonance Sensor using Automatic Nanoimprinting Lithography for Detecting LMP1 Gene |
作者: | Nai-Cheng Hou 侯乃誠 |
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: | 自動化奈米壓印技術,聚合?連鎖反應,奈米狹縫表面電漿共振晶片,免標定免疫分析,實驗室晶片, Automatic Nanoimprinting Lithography,Polymerase Chain Reaction,Nanoslit Surface Plasmon Resonance Sensor,Label-free Immunoassay,Lab on a chip, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 本研究以表面電漿共振(Surface Plasmon Resonance)晶片為基礎,發展出新的聚合酶連鎖反應(Polymerase Chain Reaction, PCR)產物分析方法。我們利用自動化奈米壓印技術(Automatic Nanoimprinting Lithography),製作奈米狹縫表面電漿共振晶片(Nanoslit Surface Plasmon Resonance Sensor),將其應用於檢測經過聚合酶連鎖反應後的產物,並藉由光譜紅移(Red-shift)現象分析實驗結果。
首先使用電子束微影製程(E-beam Lithography)、以及反應離子蝕刻技術(Reactive Ion Etching),在矽晶圓基板上定義出週期性奈米狹縫結構,再將矽晶圓基板交由廠商翻模製作鎳鈷合金材質的沖壓模具。我們以聚碳酸酯(Polycarbonate, PC)作為晶片的材料,利用自動化奈米壓印技術將沖壓模具上的結構轉印至塑膠基板上,經由遮罩以直流式真空濺鍍機(DC Sputter)對塑膠基板做局部鍍金製程。接著以黃光微影製程、以及軟微影製程(Soft Lithography),製作以聚二甲基矽氧烷(Polydimethylsiloxane, PDMS)為材料的封閉式流道系統。最後再將PDMS流道對準奈米狹縫結構,將其吸附在塑膠基板上,即完成製作奈米狹縫表面電漿共振晶片的步驟。 本研究量測的生物樣本為LMP1 Probe以及經過PCR處理後的LMP1 DNA。由於LMP1 Probe無法直接與晶片上的金鍵結,因此需要以Cysteamine對晶片表面做化學修飾才能使LMP1 Probe與晶片表面產生鍵結。接下來,再利用Probe和DNA之間的專一特性使LMP1 DNA與LMP1 Probe鍵結,並量測其穿透光譜。比較兩者的光譜,可以發現接上LMP1 Probe後會產生約1.11 nm的紅移,而接上LMP1 DNA的光譜則會產生約2.78 nm的紅移。 總結來說,我們所使用的自動化奈米壓印技術可以達到大量生產(Mass Production)、低成本(Low-cost)以及快速製程等目標,加上表面電漿共振技術具有高靈敏度(High Sensitivity)、即時(Real-time)檢測以及免標定(Label-free)的優點,且奈米狹縫表面電漿共振晶片可以精準的量測到PCR產物的紅移訊號,由此證實我們成功以表面電漿共振技術為基礎發展出新的聚合酶連鎖反應產物分析方法。 In this study, a new method of analyzing PCR product with SPR sensor has been developed. We fabricated the nanoslit SPR sensor, which is used to sense PCR product, by using automatic nanoimprinting lithography. Then we analyzed the experimental result by a red-shift of the resonant spectrum signals, which is corresponding to the numbers of probe-DNA conjugation. Firstly, the periodic nano-grating structure was clarified and fabricated on a silicon wafer. After that, we delegated the outsourcing company to transfer the structure onto a nickel-cobalt alloy mold. PC is adopted as the material of our sensor. The nanostructure was then transferred onto PC by nanoimprinting lithography. Gold was deposited on the grating structure of PC by DC sputter. Using soft lithography process, a closed microchannel system made of PDMS was produced. Lastly, we located sensors in the microchannel. This way, we completed the fabrication of nanoslit SPR sensor. In this study, LMP1 probe and LMP1 DNA processed with PCR were used as the testing samples. Owing to the fact that LMP1 probe is unable to form covalent bonds with Au, we modified Au with cysteamine. The amino-group bonds on cysteamine would form covalent bonds with LMP1 probe, and hence is able to be detected by spectrometer. Next, we utilized biospecificity of ssDNA to conjugate LMP1 probe with LMP1 DNA. By compared the resonant spectrum signals in the experiment, we found that the red-shift value of LMP1 probe is 1.11 nm and LMP1 DNA 2.78 nm. In summary, the automatic nanoimprinting lithography has several advantages such as the ability of mass production, low-cost and time efficiency, and our SPR system has the advantage of lightweight, high sensitivity, real-time detection and label-free. In addition to this, the nanoslit SPR sensor can detect the red-shift value of PCR product accurately. Through the experiment, we developed a new method of analyzing PCR product with SPR sensor. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79160 |
DOI: | 10.6342/NTU201802266 |
全文授權: | 有償授權 |
電子全文公開日期: | 2023-08-02 |
顯示於系所單位: | 應用力學研究所 |
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