利用雙重顏色氧化鋅與二氧化鈦奈米線進行癌症細胞辨識

Abstract

在現今癌症檢測技術中，最常見為螢光染色法，然而螢光染色法存在許多缺點，例如螢光衰退、螢光顏色的限制與細胞自體螢光分辨上的困難，這些都使得醫師在手術的過程中難以達成癌症細胞之即時檢測。雖然有許多團隊利用量子點並藉由表面改質的方式來解決這些困難，然而量子點在製備以及表面改質的過程不僅會改變半導體特性，並且製程昂貴。 因此，我們提出了利用氧化鋅與二氧化鈦奈米線應用於癌症細胞之辨識，並以半導體光源來代替傳統螢光染色之有機螢光物質。氧化鋅與二氧化鈦有著優異的光學特性並常應用於生醫相關研究及產品，且氧化鋅與二氧化鈦奈米線對於蛋白質皆有很高的親和力，因此我們利用具有特異性的生物抗體與奈米線的接合形成了特殊的生物標記並藉由光致發光系統以及顯微拍攝技術，從影像與頻譜的雙重鑑定，可成功辨別癌症細胞與正常細胞。 此外，我們也做了一系列的量化實驗，將生物標記的濃度、有效檢測細胞數量範圍與取得之光學頻譜分析比較並找出規律，最後更模擬真實細胞分布狀況將正常細胞與癌症細胞共同培養並且按照不同比例做癌症細胞檢測實驗，這些實驗數據與整理之規律性將可提供外科醫師在臨床手術上能夠即時辨識癌症細胞。 在這篇論文中，我們提出以半導體光源來替代有機螢光物質來改善現有螢光染色法，並利用氧化鋅與二氧化鈦奈米線與相對應抗體接合而形成之生物標記成功標識出癌症細胞與正常細胞，讓醫師能在臨床手術中進行癌症細胞之即時檢測，且量化實驗的結果也使即時檢測之技術更加系統化。

Among cancer cell detection methods, fluorescence microscopy cancer detection is one of the most common methods. However, there are many disadvantages for fluorescence cancer detection such as the phototoxicity, the limited number of available fluorescent channels, and the overlap of the excitation and emission spectra of the stains. Furthermore, under a constant light illumination, it possesses the issue of photobleaching, making real-time surgery difficult. Many groups have reported that quantum dots can conquer these problems; however, the quantum dots method requires surface modification and the surface state induced by modification changes the property of semiconductor material. Moreover, all the processes cost high. To overcome these issues, the semiconductor light sources are alternative choices. ZnO and TiO2 nanowires connected with specific antibodies are proposed to identify cancer cells and normal cells, replacing the organic fluorescent substances in the traditional cancer detection method. ZnO and TiO2 have exceptional optical properties, and they are often applied to biomedical research and commercial products. In addition, duto to great affinities between both ZnO and TiO2 nanowires and many proteins, ZnO and TiO2 nanowires bounded to specific antibodies are regarded as biomarkers to distinguish cancer cells from normal cells. Furthermore, from PL spectra and bio-images, our idea for cancer cell detection by semiconductor nanowires was confirmed. Then, a series of quantitative analyses, including biomarker concentration limit, the valid range of cell numbers, and co-culture case to simulation the real situation were conducted to examine the relationship between the optical response from biomarkers and cell numbers. The data from quantitative analyses were expected to provide a reference for surgeon for real-time cancer cell detection while performing the surgery. In this thesis, we reported a novel cancer cell detection technology by using ZnO and TiO2 nanowires connected to specific antibodies to distinguish cancer cells from normal cells, leading successful real-time cancer cell detection during cancer resection. Moreover, a series of quantitative experiments will provide physicians with key parameters and thus systemized the real-time cancer cell detection.