臺灣博碩士論文加值系統

本研究已經成功發展出結合依序釋放和分流微流體功能的離心式免疫檢測平台。於此平台中，試劑儲存的液體分流結構可大幅減低試劑注入的次數，所有檢測試劑可在檢測前預先注入到各個儲存槽，並藉由馬達控制程式使各個試劑依照檢測流程依序釋放。除此之外，免疫檢測方面利用本實驗室發展的磁盤模組操控磁珠自動的在培育步驟於混合槽中移動與在清洗步驟保留在混合槽中。最後我們將針對檢測的操作條件與液體用量進行最佳化探討，除了探討 馬達最佳化加速度和轉速使液體分流時各分流槽的誤差能降低。除此之外， 還探討最佳化的清洗液體積與呈色液體積使其完全取代偵測槽中的液體以減少偵測訊號的干擾及誤差。經過一連串的最佳化實驗後，此離心式免疫檢測平台僅須38分鐘即可自動化完成所有檢測流程且hCG的偵測極限濃度為0.15 mIU/mL。

In this work, a centrifugal microfluidic platform was developed to replace microtiter plates for conducting enzyme-linked immunosorbent assays. The sequential flow control and flow-splitting were developed to reduce the labor required for sample and reagent loading. In addition, magnetic modules were developed to conduct both incubation and washing in automation. In addition, the factors which affect the sequential flow control of the capillary valves were investigated. Finally, the centrifugal microfluidic platform was used to conduct ELISA. The assay protocol can be finished within 38 minutes with the limit of detection (for hCG) around 0.15 mIU/mL

致謝 I
摘要 II
Abstract III
目錄 IV
圖目錄 VI
表目錄 VIII
第一章 緒論 1
1.1 微流體晶片(Lab on a chip) 1
1.2 微流體光碟(Lab on a disk) 2
1.3 酵素聯結免疫吸附法(Enzyme-linked immunosorbent assy, ELISA) 3
1.4 研究動機 5
第二章、文獻回顧 8
2.1 酵素聯結免疫吸附法在微流體光碟上之應用 8
2.1.1 依序釋放微流體光碟 8
2.1.2 分流微流體光碟 12
2.2 微珠與磁珠在生物檢測上之應用 13
2.3 磁性粒子之操控 15
2.4 相關微流體功能在酵素聯結免疫法檢測之應用 19
2.3 生物標記技術之應用 20
第三章、實驗設計 25
3.1 實驗藥品與操作儀器 25
3.1.1 ELISA檢測藥劑 25
3.1.2 免疫性磁性奈米粒子製備 25
3.1.3 微流體光碟製作材料 26
3.1.4實驗儀器 26
3.2微流體光碟製作 27
3.3實驗操作流程 28
3.3.1磁珠表面修飾人類絨毛膜性腺激素抗體 28
3.3.2 酵素連結免疫吸附法操作流程 30
3.4實驗檢測儀器 35
3.4.1 光學式檢測儀器 35
3.4.2馬達操作系統 36
3.4.3 影像拍攝分析系統 37
第四章、結果與討論 39
4.1 微流體功能探討 39
4.1.1微流閥功能探討 39
4.1.2 加速度與轉速對液體分配之探討 42
4.1.3 液體取代 47
4.2 光碟式酵素連結免疫吸附法最佳化操作條件 49
4.2.1 磁珠表面修飾人類絨毛膜性腺激素抗體之複合物活性檢測 49
4.3 光碟操作條件最佳化 51
4.3.1 高低水位培育最佳化之探討 51
4.3.2 磁珠混合運動最佳化 52
4.3.3 清洗液體積最佳化 54
4.4 微滴定盤ELISA與光碟式ELISA檢測結果比較 55
第五章、結論 59
第六章、未來展望 60
第七章、文獻參考 61

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