研究生: |
廖登正 Liao, Teng-Cheng |
---|---|
論文名稱: |
利用表面增強拉曼光譜技術檢測口腔癌細胞之研究 Detection of oral cancer by Surface Enhanced Raman Spectroscopy |
指導教授: |
柳克強
簡世森 |
口試委員: |
林滄浪
黃穰基 柳克強 簡世森 |
學位類別: |
碩士 Master |
系所名稱: |
原子科學院 - 工程與系統科學系 Department of Engineering and System Science |
論文出版年: | 2015 |
畢業學年度: | 103 |
語文別: | 中文 |
論文頁數: | 94 |
中文關鍵詞: | 拉曼光譜 、表面增強拉曼散射 、口腔癌 、膜表面蛋白 |
相關次數: | 點閱:3 下載:0 |
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本研究目的在發展拉曼光譜儀作為診斷口腔癌細胞的工具。口腔癌是國人男性罹患率很高的疾病,治療困難、死亡率高的疾病。目前台灣口腔癌是男性腫瘤死亡率第五名,平均減壽約十六年。特定膜蛋白的表達已被證實與口腔癌有關,位於細胞外圍的「膜蛋白體」扮演細胞發生癌變的關鍵角色,但普通的檢測方法,例如: 病理組織診斷(Histopathological)免疫化學法(Immunochemical assay)和免疫印記法(Western blot)不足以立即檢測,且需要專業病理學者做正確判斷。拉曼光譜(Raman spectra)可以提供即時性、非侵入性,目前被廣泛使用來鑑別診斷正常、癌前期和癌細胞。表面增強拉曼光譜是一個有效偵測口腔細胞的方法,我們希望藉由此研究建立與口腔癌有關的表面蛋白之拉曼光譜資料庫(data base),作為將來使用拉曼光譜技術診斷口腔癌之用。
由於蛋白質之拉曼訊號微弱,所以選擇適當的拉曼活化基板是非常重要的。我們將利用中研院王玉麟教授團隊所開發之表面增強拉曼活化基板(SERS-active substrate)來增加表面蛋白的拉曼訊號,此表面增強拉曼活化基板為奈米銀陣列所製。我們研究將現成之純化表面蛋白樣品施加於此基板之方法,使表面蛋白透過共價鍵鍵結或是靜電力(electrostatic force)附著於基板上,並確認此基板對表面蛋白有增強拉曼之效果,以建立表面蛋白之拉曼光譜資料。我們使用戊二醛來固定表面增強拉曼活化基板上的細胞,保持細胞完整性且不破壞細胞壁。口腔細胞的表面拉曼光譜效應,可在很短的採集光譜時間被觀察到,進而可獲得口腔癌的表面蛋白拉曼光譜。
我們建立了三種不同的口腔癌(OCSL、SAS 、FADU)及兩種不同的正常細胞(PUP-34、Fibroblast)其拉曼光譜資料庫。從拉曼光譜的特徵峰,可以區別出五種不同的口腔細胞。我們將口腔細胞做特徵峰相關性處理,從口腔細胞特徵峰相關係數獲得標記辨識率。最後在表面增強拉曼活化基板上塗佈膠原蛋白(Collagen),藉由膠原蛋白吸附口腔癌細胞(SAS),觀察到更穩定的拉曼特徵峰。牛血清白蛋白(BSA)修復SAS細胞可獲得較原始的細胞,修復後的SAS細胞拉曼重現性不好。所以表面增強拉曼活化基板是一個快速且有效診斷口腔癌細胞拉曼的方法。實驗完成後可以確定建立的拉曼光譜系統可以應用到臨床上,以增加口腔癌治療率,造福國人。
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