- 學位論文
整合預濃縮微米流道與表面電漿共振之奈米狹縫生物晶片用於高靈敏度免標定免疫分析檢測
Integration of Preconcentration micro-channel with Nanoslit Surface Plasmon Resonance Chip for high sensitivity Immunoassay detection
摘要
本研究的目標在於開發將表面電漿共振晶片與預濃縮器結合之新型免標定免疫分析生醫晶片，藉由熱轉印技術製作奈米狹縫(Nanoslit)表面電漿共振晶片(Surface Plasmon Resonance,SPR)，再利用聚二甲基矽氧烷(PDSM)製作預濃縮微米流道，配合Nafion奈米孔隙流道做為離子選擇性通道，結合成一個新型的生醫檢測平台。將活性較高的分子修飾在共振檢測金屬薄膜上，藉由電壓調整控制濃縮區塊於檢測共振金屬上方，結由活性分子與待測生物檢體結合，產生光譜訊號紅移進行免疫分析。 本實驗利用射出成型技術製作表面電漿共振晶片，取代熱壓印技術，提升鏡片製作的效率並降低製作成本，再利用矽晶圓塗佈SU-8光阻，將流道藉由微影技術(Lithography)成型矽晶圓上製作出可重複利用之母模，再利用熱蒸鍍機將鋁金屬薄膜鍍上共振奈米狹縫，接著使其金屬表面氧化，並利用3-氨基丙基三乙氧基矽烷(3-Aminopropyltriethoxysilane, APTES)，使檢測區表面官能化，以利與待測樣本結合。流道設計上配合施加電壓，設定出最佳的空乏區長度(Ion Depletion Zone)與濃縮倍率，將低於檢測極限之樣本透過預濃縮技術聚集於檢測區域中，提升檢測最低極限。 本實驗使用之樣本為人類免疫球蛋白G之抗原 (Immunoglobulin G)、抗體(Anti-Human IgG)，將表面修飾帶有胺基之共振晶片通入高濃度100 ug/mL人類免疫球蛋白G(IgG)，光譜儀檢測極限為1ug/mL Anti-IgG，依序通入低於極限之樣本最後成功將量測極限提高至1 ng/mL Anti-IgG，並且濃縮倍率介於1000~10000倍之間。
並列摘要
In this thesis, we integrated nanoslit surface plasmon resonance sensor and microfluidic preconcentration onto a chip. With the Injection Molding technique to formulate periodic grating as nanoslit and to fabricate the micro channel with Polydimethylsiloxane (PDMS), we choose Nafion 117 as ion-selective nanochannel and use dual-loop electric current to control the ion-depletion zone, maintaining the ion-enhancement region on the sensor for real-time detection. The microchannel’s mold was made on silicon wafer by Lithography technology. Using thermal evaporator deposited aluminum thin film plated on SPR sensor, and deposited Aluminium Oxide by atomic layer deposited (ALD), modified the surface by 3-Aminopropyltriethoxysilane (APTES). The samples of Immunoglobulin G and anti-human IgG were used in this experiment. the Human Immunoglobulin G(IgG) with a high concentration 100 ug/mL was transfused into the microchannel bonding with the surface modified with amine group. Furthermore, enhance the concentrations below the limit od detection of anti-IgG by ion concentration polarization (ICP), and the limit of detection (LOD) was successfully increased to 1 ng/mL anti-IgG, and the concentration rate was between 1000-10000 times. We have demonstrated a novel, label-free, high-sensitivity, real-time, portable device.