超高靈敏表面電漿螢光微脂囊陣列型測試晶片---奈米粒子耦合晶片開發/超高靈敏度表面電漿晶片與偵測系統---總計畫及子計畫一(I); Ultra-High Sensitive Surface Plasmon-Coupled Fluorescence Lipsome Multiple Spots Sensor Chip (I)

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题名:	超高靈敏表面電漿螢光微脂囊陣列型測試晶片---奈米粒子耦合晶片開發/超高靈敏度表面電漿晶片與偵測系統---總計畫及子計畫一(I);Ultra-High Sensitive Surface Plasmon-Coupled Fluorescence Lipsome Multiple Spots Sensor Chip (I)
作者:	陳啟昌;周晟
贡献者:	光電科學與工程學系
关键词:	醫學工程
日期:	2008-07-01
上传时间:	2010-12-28 15:32:10 (UTC+8)
出版者:	行政院國家科學委員會
摘要:	本研究計劃提出一種創新的表面電漿波激發螢光微酯囊平面波導微陣列以及方法及量測技術，它可即時量測生物分子間交互作用並具有近場量測能力。完成微陣列元件製作為本研究計劃的目的，建立蛋白質動力學上的即時測量平台技術。可應用於開發臨床藥物過程中若干關鍵性數據的提供。研究初期我們先著眼在一維的 Fiber-Optical Biosensor 上，我們利用光纖之消逝波激發奈米金屬粒子的表面電漿，再利用此激發螢光，待此項技術純熟且靈敏度夠高時，再推廣至二維的平面波導上。另一方面，本計畫擬結合微脂囊 (liposome) 技術，借重其獨特的訊號放大效果，應用於微陣列晶片的研發。研究初期，以發展測量血液中 LDL (low density lipoprotein) 含量之微脂體型免疫化學檢驗技術為首要目標。微脂囊檢驗法 (LIA: liposome immunoassay) 對測量花生中之黃麴毒素 (aflatoxins) 以及玉米中伏馬鐮孢毒素 (fumonisins) 的可行性已在過去實驗、發表的論文中被確定。這些檢驗法的理論基礎乃建立在被檢測物與其抗體間特異專有的免疫性辨識結合作用(immunorecognition reaction)，並利用包有訊號物質之微脂囊 (marker-entrapped liposomes) 當作訊息放大劑 (signal amplifier)。不同於酵素免疫檢驗法 (ELISA:需要再更換試劑並耗時等待酵素與其基質作用改變顏色以產訊息)，微脂囊之破裂而釋放出的光學或電學訊號提供了」立即式」的訊號放大作用。不但簡化傳統的檢測步驟以減少勞力的投資；又由於微脂囊的製程簡單，價錢不如酵素昂貴，因而降低整組試劑的成本。若將微脂囊檢驗法應用於微陣列晶片的研發上，則可改善其分析速度 (high throughput and turnover rate)。加速並加強監測心血管相關疾病病患的生理情況，以期有效控制病情。在光波導的設計與製作多點偵測晶片方面，我們將利用積體光學常用之 Beam Propagation Method 與Finite-difference time-domain 計算多模波導的厚度與消逝波的關係，並計算波導的 optical field 分佈的情形，以使得生物晶片之表面消逝波與奈米微粒產生耦合，引發更高的螢光效果。製作方面，我們將利用矽為基材（如SiO2 或SiNx）的薄膜在矽晶片上製作波導，並將 fiber biosensor 與liposome 的偵測技術建立於波導上，成為多點的測試晶片，以提高晶片的測試的同時性。本計畫的最終目的是要結合面表面電漿波激發螢光與微脂囊兩技術的平面波導微陣列，除了能將信號偵測濃度帶向更低的極限，也可以陣列的方式使檢驗的同時性增加，提高其應用性。我們也將使用台北市立聯合醫院所提供之樣本測試研究成果的實用性。 We propose to develop a novel surface plasmon-coupled fluorescence (SPCF) based fiber-optic and planar wave guide protein affinity sensor chip where the fluorescence signal is excited by surface plasma wave (SPW) on gold nanoparticles. The advantages of SPCF biosensor not only present its higher excited efficiency of the fluorescence but also the specific direction of the fluorescence emission. They are able to enhance the detection sensitivity on fluorescence detection much higher than conventional total internal reflection (TIR) method. In addition, the localized enhancement of the electric field of the evanescent wave (EW) near gold nanoparticles strengthens SPCF simultaneously. Therefore, to combine higher sensitivity of fluorescence detection with the localized properties of EW and SPW are able to real time monitor the biomolecules interaction in the near-field region simultaneously. The association rate constant (Ka) and the dissociation rate constant (Kd) of binding kinetics of biomolecules and also the equalization classicization sate constant (KD = Ka / Kd) are obtained at the same time. In addition, a liposomal array immunosensor for the measurement of the multiplex cancer markers in blood will be developed in the proposed study. Development of the liposomal immunobiosensing devices for biologically important molecules in human blood (such as insulin, AFP, CEA, β-hCG, or PSA), pathogens and mycotoxins in foods have been reported previously by Ho』s group. This type of sensors provides higher sensitivity, lower cost, speed and simplicity, which is the ultimate goal of the proposed study. The hyphenation of liposomal immunoassays and array sensor allows the quantitation of analyte-of-interest (various cancer markers in this case) with better turnover rate and improved throughput. In practice, this array sensor will help in facilitating the monitoring and management of physical status for cancer patients, and that eventually has potential as a rapid and inexpensive diagnostic assay used either at home care or clinics. For designing the optical planar waveguides to form multiple spots sensor chips (MSSC), beam propagation method will be used. The evanescent wave propagating on the surface of the multimode waveguides will be studied by varying the thickness of the waveguides. This study will help to increase the coupling of the evanescent wave into the SPW on the surface of gold nanoparticles within the evanescent field of waveguide. For realizing the protein affinity sensor chip, the multi-mode planar waveguides will be fabricated in silica (SiO2) materials on silicon substrates. With the precise semiconductor photolithography fabrication techniques, the thickness of the waveguides can be determined and the evanescent waves can be well controlled. We will combine the surface plasmon-coupled fluorescence and the liposomal immunobiosensing with the optical plane waveguide to increase the throughput of the detection. With this SPCF and liposomal MSSC, one can study and measuring the kinetics behavior of protein-protein in general. The final aim of the project is to combine the surface plasmon-coupled fluorescence and liposomal array immunosensor methods on an array chip. By putting the gold particles in the liposomes, we plan to boost the detection sensitivity to higher level. By using the array chip, the throughput will be enhanced. The devices will also be used to do clinical test with the whole blood samples of patients provided from the Taipei City Hospital. 研究期間：9608 ~ 9707
關聯:	財團法人國家實驗研究院科技政策研究與資訊中心
显示于类别:	[光電科學與工程學系] 研究計畫

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