- 學位論文
結合微流道之銦鎵鋅氧化物薄膜電晶體應用於蛋白質與生物素混合反應之檢測
Investigation of Protein-Biotin Interactions by IGZO Thin Film Transistors Integrated with Microfluidic Channels
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摘要
在這篇論文中介紹以氧化銦鎵鋅薄膜電晶體與感測金屬板構成之生醫感測器檢測生物分子反應的混合狀態，此研究分兩部分： 在第一部分，我們將以薄膜電晶體生物感測器與外接的微流道系統結合，應用於偵測生物素與鏈親和素的混合反應與流體擴散性質。由於濃度梯度的關係，待測物溶液會經由微流道流至金感應板而被生物感測器偵測到。我們首先分別量測生物素與鏈親和素的訊號，得到擴散性質與電流的關係。之後將生物素與鏈親和素混合並量測其隨時間變化的電流性質，以探究兩者之間的反應行為。此外也進行螢光染色的影像拍攝去驗證蛋白質複合物間的結合。 在第二部分，為了進一步分析生物素與鏈親和素的混合情況，我們改用Y型混合器流道。此外，使用聚二甲基矽氧烷來密封微流道系統以避免待測物溶液的蒸發。接著先分別量測生物素與鏈親和素的電流訊號，以定義待測物的擴散時間。然後進行一系列同時與時間差的混合實驗。藉由觀察即時的電流變化，來分析待測物在流道中的混合狀況。最後我們使用牛血清白蛋白做為對照組，以驗證薄膜電晶體生醫感測器之非特異性結合的情況。
並列摘要
In this thesis, a biosensor consists of an Indium-Gallium-Zinc-Oxide (IGZO) thin film transistor (TFT) biosensor and a gold sensing pad is demonstrated for the detection on biomolecule’s reactions. There are two parts in this thesis. In the first part, a TFT biosensor with microfluidic channels is demonstrated for the detection of biotin and streptavidin reaction and their diffusion behaviors. Due to the concentration gradient, the analyte solutions may flow directly through the microfluidic channels to the sensing area and sensed by the biosensors. The diffusion experiments of streptavidin and biotin are first conducted separately to understand the diffusion properties. Then the binding interaction of streptavidin-biotin complexes is detected and analyzed in time domain. We also verify the binding among streptavidin and biotin by using the fluorescence microscope. In the second part, to further analyze the mixture condition of streptavidin and biotin, a Y-type external microfluidic channel is demonstrated. In addition, the channels are sealed by polydimethylsiloxane to avoid evaporation of the target analyte solutions. Next, the current signals of streptavidin and Biotin are measured to define the diffusion time. Then, a series of mixture and time delay experiments are conducted. By observing the real-time current change of the TFT biosensor, the mixture condition of the reaction can be analyzed. Finally, bovine serum albumin (BSA) is used for a control experiment to verify the specificity and reliability.
參考文獻
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