本研究利用化學氣相沉積法 (Chemical Vapor Deposition, CVD)製備大面積石墨烯， 並用濕式轉印法疊加雙層石墨烯至二氧化矽基板，製作平面閘極石墨烯場效電晶體 微架 構的 生醫感測器，接著利用低損傷 氮 電漿對石墨烯進行共價功能化 改質 ，以利後續接上 抗體 實現 鼻咽癌循環腫瘤細胞的 無標記檢測 並 透過 電晶體的 電性 上 之 狄拉克點的訊 號變化 量來判斷檢測物之濃度。其優勢為相較於醫學上的檢測方法如 :核磁共振、電腦斷 層與 聚合酶連鎖反應 石墨烯場效電晶體生醫感測器在感測時間、單位晶片成本與儀器 成本都更低。 本研究選擇鼻咽癌循環腫瘤細胞作為感測標的物，利用其細胞膜上的上表 皮細胞黏附分子進行細胞的捕捉與感測。 研究結果 顯示 低損傷氮電漿成功的使石墨烯表面胺化，後續進行的 X射線光電子能 譜 分析顯示氮的原子比 由原本的 1.07%提升到 且分峰後的結果顯示氮鍵結組成 中的 C-NH2鍵 約占 並以螢光顯微鏡初步對胺化石墨烯分析，結果顯示胺化石墨 烯接上的抗 體數量大於普通石墨烯。本研究所製作的 感測器 的 線性檢測範圍為 1 cell/15 l到 1000 cell/15 l 檢測極限 (LoD)為 1 cell/15 l 並且其靈敏度可達 -0.040 V /decade 線性回歸度 (R2)達 0.93，並且在後續的 專一性 測試中表現出了高度的 專一性 。同時也利 用電子顯微鏡觀察捕捉細胞後的石墨烯表面，並計算出其捕捉密度為 238.6 ± 29.8 cell/mm2。 本研究可提供未來 針對鼻咽癌循環腫瘤細胞 快速且 高可靠度之 精準檢 測，可 用來作為鼻咽癌早期發現與治療後後續追蹤的平台。;In this study, a planar gate graphene field effect transistor biomedical sensor was fabricated by using large area CVD graphene and transferred to SiO2 substrate by wet etching method. Graphene is covalently functionalized by the low damage plasma to facilitate subsequent binding of antibodies to achieve label-free detection of circulating tumor cells of nasopharyngeal carcinoma. Compared with more conventional detection methods such as magnetic resonance imaging (MRI), computed tomography (CT) and polymerase chain reaction (PCR), the graphene field effect transistor biomedical sensor is lower in unit cost, instrument cost and the sensing time is shorter. Circulating tumor cells(CTC) is selected as the target. Epidermal cell adhesion molecules on the cell membrane were used in the capturing and sensing of CTC The results showed that the low-damage nitrogen plasma successfully functionalize the graphene surface. Subsequent X-ray photoelectron spectroscopy (XPS) analysis showed that the atomic ratio of nitrogen increased from 1.07% to 3.17%, and the results after fitting showed that com position of C-NH2 bond in the nitrogen related bond is about 62.8%. Fluorescence microscope shows that the number of antibodies attached to the functionalized graphene is greater than that of pristine graphene. In testing, our biosensors revealed high selectivity and limit of detection receive 1 cell/15 l, the detection linear range at 1 cell/15 l to 1000 cell/15 l (R2=0.93) and sensitivity can reach -0.040 V /decade, shows high reliability and characteristic. Moreover, we used the scanning electron microscope (SEM) to observe the surface of graphene after capturing of CTC and the capture rate was at 238.6±29.8 cell/mm2. Our biosensors accurately detect nasopharyngeal carcinoma circular tumor cell rapidly and highly reliable, and can be used as a platform for early detection of nasopharyngeal carcinoma and post treatment monitoring.
