Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/1640
標題: 利用微奈米環境提升免疫檢測效果
Enhancing sensitivities of immunoassays by nanoenvironment
作者: 陳彥廷
Chen, Yen-Ting
關鍵字: 3D gold nano particle seneor;3D奈米金顆粒感測器;serpentine microfludic channel;Der p2;S型微流裝置;歐洲式塵螨
出版社: 機械工程學系所
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摘要: 
本研究成功利用3D奈米金顆粒感測器搭配微流裝置來偵測塵螨過敏原(Derp2)。首先利用陽極氧化製程製作陽極氧化鋁模(AAO),接著蝕刻陽極氧化鋁模,使得背阻障層呈半球形狀。利用電鑄技術,將Ni沉積上製成3D凹洞狀Ni模。接著使用熱壓技術製成3D半球型PC塑膠材料。接著在3D半球形PC上濺鍍一層金導電層,並利用電化學沉積法於奈米結構上均勻沉積大小約為10 nm之奈米金顆粒,利用此技術提升感測區域面積,結合微流環境形成高靈敏度奈米生醫檢測晶片
,進行塵螨相關環境偵測。最後利用螢光檢測以及阻抗量測進行環境塵螨濃度的檢測,並發現此感測晶片利用浸泡法,在螢光檢測中最低檢測濃度可達10pg/ml並擁有11.44%螢光覆蓋率,而阻抗量測最低檢測濃度可達1pg/ml;當加入微流系統進行接附時,在螢光檢測中最低檢測濃度可達0.1pg/ml並擁有13.9%螢光覆蓋率,而阻抗量測最低檢測濃度可達0.1pg/ml,且螢光檢測會因感測區域接近飽和而導致螢光覆蓋率提升不明顯;阻抗量測在高濃度時仍呈線性關係,因此阻抗量測系統所能量測範圍大於螢光檢測。

A 3D gold nano particle (NP) seneor in serpentine microfludic channel (S channel) was developed and has the advantage of label-free detection, high sensitivity and low cost. In the fabrication of 3D gold NP sensor, we demonstrated a replica mold fabrication method for nano hemisphere arrays by nanomolding nickel electroforming the barnier-layer surface of an anodic aluminum oxide (AAO) as the master mold. The fabricated 3D nickel mold is further used for replica molding of a nano-structure polycarbonate (PC) substrate by hot embossing. Afterwards, a layer of gold film and 3D NPs were deposited on the PC substrate by sputtering and electrochemical deposition, respectively. Compared to a flat gold film, 3D gold NPs have 3 times surface area which can highly increase the sensitivity of sensor. Furthermore, a serpentine microfludic channel was fabricated by soft lithography and polydimethylsiloxane (PDMS). Then, 3D gold NPs sensor were bonded on the S channel by O2 plasma treatment. D. pteronyssinus (Der p2) and specific antibody (C1) were used to present the performance of sensor. Electrochemical impedance spectroscopy (EIS) and fluorescence detection by confocal microscope were used to detect the concentration of Der p2.
In the EIS analysis, the minimum detectable concentrations of Der p2 were down to 0.1pg/ml and 10pg/ml when the 3D gold NPs sensor in S channel and in immerse deposition, respectively. In the fluorescence detection, the average coverages were 35.59% and 11.44% at the concentration of Der p2 10pg/ml, when sensor in S channel and in immerse deposition. Experiment at results showed that 3D gold NP sensor in S channel have 100 times sensitivity than sensor than sensor in immerse deposition. In the other hand, EIS analysis showed larger sensing linearity than fluorescence detection because fluorescence was saturated when the concentration of Der p2 was larger than 1ng/ml. Our results showed that 3D gold NPs sensor in S channel have great sensitivity (0.1pg/ml) compared to ELISA (1ng/ml).
URI: http://hdl.handle.net/11455/1640
其他識別: U0005-2208201115352200
Appears in Collections:機械工程學系所

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