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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/30675

Title: 開發抗登革病毒與腸病毒71型的藥物及疫苗之快速篩選-細胞包覆耗氧感測微液珠陣列晶片應用於篩選抗登革病毒與腸病毒71型感染之藥物(子計畫二)( III )
Authors: 黃士豪
Contributors: NTOU:Department of Mechanical and Mechatronic Engineering
國立臺灣海洋大學:機械與機電工程學系
Keywords: 光學;氧氣感測;相位量測;登革熱病毒
Phosphorescence;OCR;Phase detection system;Pneumatic sealed activation device
Date: 2011-08
Issue Date: 2012-04-13T01:18:31Z
Abstract: 摘要:本研究目的將整合細胞培養微陣列晶片、氣動密閉作動裝置、與磷光相位差光學檢測系統,設計一微小化、自動化和陣列化之微陣列生物晶片,透過投影機內的Digital Micromirror Device (DMD),可控制UV-LED的光源投影圖形,激發欲檢測之區域的耗氧量,經Labview程式,達到自動化的即時激發量測效果。本研究之微型生物晶片裝置暨檢測系統,主要分三個部份:(1)細胞培養微陣列晶片:此晶片藉由微機電的製程技術,在玻璃基材上製作出微陣列孔洞晶片,將氧氣感測螢光薄膜製作於孔洞底部,並將細胞培養貼附於微陣列孔洞內,可進行長時間細胞培養。(2)氣動密閉作動裝置:藉由微機電的製程技術製作此氣動密閉作動裝置,具有可氣壓上下致動的閥座密封上述細胞培養微陣列晶片中的微陣列孔洞,藉以降低外界氧氣滲透問題。(3)磷光相位差光學檢測系統:此光學系統包含數位微反射鏡裝置,可控制激發光的光源與投影圖像,激發上述上述細胞培養微陣列晶片中的微陣列孔洞內的氧氣感測螢光薄膜,達到自動化的即時激發量測效果,並透過磷光相位差計算處理,藉此得知孔洞內的細胞耗氧量(oxygen consumption rate, OCR)。上述微型生物晶片裝置暨檢測系統已成功應用於細胞的耗氧率定量量測、病毒感染後細胞耗氧率之定量量測、以及病毒抑制藥物對病毒的抑制性之初步量測上。本裝置藉由微機電的製程技術,在玻璃基材上製作直徑為1mm之微陣列孔洞晶片,將氧氣感測螢光薄膜製作於孔洞底部,並將細胞培養貼附於微陣列孔洞內,利用氣壓閥的作動密封微陣列孔洞晶片來降低氧氣滲透問題,並藉由耗氧率校正公式修正在半密閉式系統中所造成的氧擴散問題,已成功完成定量細胞(1×103/per well)下的耗氧率、並驗證其重複性及穩定性;及細胞於M.O.I=0.1、0.5、1的病毒液濃度下的耗氧率(OCR)代謝情況,由實驗耗氧率(OCR)的量測結果可即時得知在感染後的第9個小時,M.O.I=1的存活率為0,而M.O.I=0.1、0.5的存活率分別60%、40%。藉由耗氧率(OCR)的量測結果更進一步量得登革病毒對抗體及藥物的抑制,藉由將對登革病毒病毒具療效之專一抗體注入晶片內,可於數小時內驗證抗體確有治病之療效,免除傳統免疫分析之繁瑣與費時,亦無須藉由複雜之動物實驗驗證,初步達成確認疫苗抗原研究目的。同時配合子計畫一所提供之藻類萃取物-鹿藻膠,驗證其確有抵抗登革熱病毒之能力。藉由本研究所提出的微型檢測系統,可即時時動態量測病毒感染後細胞耗氧率之定量量測、以及病毒抑制藥物對病毒的抑制性之初步量測上。此創作便於檢測研發新興藥物或疫苗,了解細胞毒化後每個階段的代謝情況,進而達到藥物快速篩選、疾病檢測、與疫苗開發之目的。
abstract:The main objective of this study is to integrate cell culture micro array chip, pneumatic sealed activation device and phosphorescence phase difference optical detection system so as to design a miniaturized, automatic and array micro array biochip. Through Digital Micromirror Device (DMD) in the projector, the projection figure of UV-LED optical source can then be controlled, and the oxygen consumption in the area to be detected can then be excited, then through Labview program, the automatic and real time excitation measurement effect can then be achieved. The micro biochip device and detection system of this study can be mainly divided into three parts: (1) Cell culture micro array chip: For this chip, MEMS (Micro Electro Mechanical Systems) process technology is used to prepare micro hole array chip is prepared on glass substrate, and oxygen sensing fluorescence thin film is fabricated on the bottom of the holes, meanwhile, cell culture is attached within the array holes to perform long time cell culture. (2) Pneumatic sealed activation device: Through the MEMS process technology, this pneumatic sealed activation device is prepared, and it has pneumatically up-and-down activated valve seat to seal the micro array holes in the above cell culture micro array chip so as to reduce the external oxygen penetration issue. (3) Phosphorescence phase difference optical detection system: This optical system includes Digital Micromirror Device to control the optical source and projection image of the excitation light. It excites the oxygen sensing fluorescence thin film within the micro array hole of the above cell culture micro array chip so as to achieve automatic and real time excitation measurement effect. Meanwhile, through phosphorescence phase difference calculation, the cell oxygen consumption within the hole can then be obtained. In the experiment, cell oxygen consumption rate has been successfully measured quantitatively, meanwhile, the oxygen metabolic situation of the cell under different M.O.I has been repeatedly tested, furthermore, the inhibition on virus from specific antibody and medicine has been further measured. This innovation facilitates the detection and development of newly emergent medicine or vaccine, the understanding of metabolic stage at each stage after cell intoxication, finally, the objectives of fast medicine sorting, disease detection and vaccine development can then be achieved.
Relation: NSC100-2627-B019-002
NSC100-2627-B019-002
URI: http://ntour.ntou.edu.tw/handle/987654321/30675
Appears in Collections:[機械與機電工程學系] 研究計畫

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