矽多層次奈米結構應用於捕捉鼻咽癌循環腫瘤細胞與EB病毒DNA偵測

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陳奕瑋(Yi-Wei Chen) 查詢紙本館藏

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材料科學與工程研究所

論文名稱

矽多層次奈米結構應用於捕捉鼻咽癌循環腫瘤細胞與EB病毒DNA偵測
(Si Hierarchical Nanostructures for Capturing Nasopharyngeal Cancer Circulating Tumor Cells and Detecting EBV DNA)

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摘要(中)

藉由研究循環腫瘤細胞(circulating tumor cells, CTCs)可以幫助我們了解癌細胞轉移的生物學，透過監測患者血液中CTCs的數量更可以幫助我們了解治療的方式是否有效。大多數癌症病患的死因是由轉移的癌細胞導致，而非原發腫瘤，因此若能於癌症初期就從血液中偵測到CTC，將提供早期的診斷或甚至於個人化的治療。
本實驗利用一種低成本的濕式蝕刻法製備出矽多層次奈米結構 (Pyramid/Nanowire, NW)來捕捉血液中的鼻咽癌(nasopharyngeal cancer, NPC)細胞。根據實驗結果顯示，金字塔結構可以透過它的幾何形狀來增加表面積，使基板表面上抗體的數量增加，因能夠提升細胞與基板產生鍵結的機會。此外，奈米線結構可以增加細胞與基板之間的附著性，因為癌細胞表面的偽足可以延伸至奈米線與奈米線之間的縫隙中，使Pyramid NW結構成為一個理想的CTC捕捉基板。
大多數鼻咽癌病患血液中EBV DNA的濃度會高於一般人。近年來由於表面增強拉曼散射(SERS)具高靈敏度來偵測分子的特異性，這項技術已可用來偵測DNA。本實驗中僅需簡單的將Pyramid NW基板泡入硝酸銀溶液中，即可變成一個良好的SERS基板，其DNA偵測的極限可達到10-13M，增強因子經由計算後也高達107。

摘要(英)

Studying circulating tumor cells (CTCs) helps us understand the biology of metastasis. Monitoring CTCs counts in patients’ blood can also help us know if the treatment works or not. Most cancer patients were died because of distant metastases instead of primary tumor. Thus, early detection of CTCs could provide early diagnosis of metastases and even personalized therapy.
Herein, we introduced a new CTC capturing substrate composed of Si hierarchical nanostructures (Pyramid NW) method to capture nasopharyngeal carcinoma (NPC) cell in vitro. According to the results, the pyramid microstructure increased the surface area because of its geometric shape. The amount of antibodies on the substrate also increase, enhancing the cell-binding ability of the substrate. The nanowire (NW) structure improved the cell adhesion by letting the cytoskeleton of the cells extend into the gaps between the NW, making pyramid NW structure an ideal substrate for CTC capture.
The Epstein-Barr virus (EBV) DNA titers in NPC patient’ blood is usually higher than normal people. In recent years, Surface enhanced Raman scattering (SERS) has become an efficient technique for detecting DNA because of its single molecule level sensitivity and molecular specificity. In this study, our pyramid NW substrate can easily be transformed into a SERS substrate by dipping it into AgNO3 solution. The DNA detection limit can reach up to 10-13M and the calculated enhancement factor can achieve up to 107.

關鍵字(中)

★ 循環腫瘤細胞循環腫瘤細胞循環腫瘤細胞
★ 鼻咽癌
★ 金字塔/奈米線
★ EBV DNA
★ 表面增強拉曼散射

關鍵字(英)

★ CTCs
★ NPC
★ Pyramid NW
★ EBV DNA
★ SERS

論文目次

摘要 I
ABSTRACT II
致謝 III
目錄 IV
圖目錄 VII
表目錄 IX
第一章文獻回顧 1
1.1 循環腫瘤細胞 1
1.2 捕捉CTCS的方法 2
1.3 金字塔/奈米線結構之製備 4
1.3.1 金字塔蝕刻機制 4
1.3.2 矽奈米線蝕刻機制 5
1.4 拉曼光譜 8
1.4.1 拉曼散射原理 8
1.4.2 表面增強拉曼散射 9
1.5 鼻咽癌簡介 12
1.6 接觸角之相關理論 13
1.6.1 Young’s Equation 13
1.6.2 Wenzel model & Cassie model 13
1.7 研究動機 16
第二章實驗方法 19
2.1 實驗流程 19
2.2 實驗藥品 20
2.3 分析儀器 21
2.3.1 掃描式電子顯微鏡 (Scanning Electron Microscopy, SEM) 21
2.3.2 螢光顯微鏡 (Fluorescence Microscope) 21
2.3.3 顯微拉曼光譜儀 (Micro-Raman Spectrometer) 22
2.3.4 接觸角量測儀 (Contact Angle) 22
第三章利用矽多層次奈米結構於捕捉癌細胞 23
3.1 實驗步驟 23
3.1.1 製備金字塔/矽奈米線結構 23
3.1.2 表面抗體修飾處理 23
3.2.3 癌細胞培養與偵測 24
3.2 結果與討論 25
3.2.1 基板結構與形貌分析 25
3.2.2 細胞數量檢測 28
3.2.3 細胞形貌分析 31
3.3 結論 34
第四章利用矽多層次奈米結構之表面增強拉曼散射基板於EBV DNA感測 35
4.1 實驗步驟 35
4.1.1 製備金字塔/矽奈米線結構 35
4.1.2 表面修飾奈米銀顆粒處理 35
4.1.3 DNA鍵結 35
4.2 結果與討論 38
4.2.1 基板結構與形貌分析 38
4.2.2 表面增強拉曼散射強度分析 40
4.2.3 接觸角分析 47
4.3 結論 49
參考文獻 50

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指導教授

李勝偉(Sheng-Wei Lee)

審核日期

2017-7-13

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