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研究生:林佾鴻
研究生(外文):Yi-Hung Lin
論文名稱:具有連續進樣裝置之微流體電泳晶片之研發
論文名稱(外文):Flow-through Sampling for Electrophoresis-based Microfluidic Chips Using Hydrodynamic Pumping
指導教授:陳淑慧陳淑慧引用關係
指導教授(外文):Shu-Hui Chen
學位類別:碩士
校院名稱:國立成功大學
系所名稱:化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:126
中文關鍵詞:連續進樣電泳晶片
外文關鍵詞:Flow-throughMicrofluidic Chips
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本論文是發展一套連續壓力流進樣偶合晶片式電泳分析的技術。晶片的製作是利用微機電製程的技術在石英和玻璃的基材上形成微管道。在進樣方面,樣品是經由外接的管路,以壓力流的方式導入至晶片的進樣端,當樣品被壓力流帶入至晶片中,隨即會根據晶片內微管道的流阻,進行分流。當樣品分流至晶片下端的微管道交叉處,會受到抑制電壓的影響,經由抑制電壓的關閉或開啟和可決定樣品的注入和分析,而所需施加的抑制電壓大小必須達到臨界抑制電壓值之上,此時的抑制電壓才有閘門的功能,可使分析樣品不受下一時間點的樣品干擾。在臨界抑制電壓的決定上,它與晶片幾何形狀,連續壓力流大小,電泳緩衝溶液的特性有關。在應用方面,利用壓力進樣所得到的BSA和Anti-BSA的結合常數為3.3*107M-1與文獻利用電動進樣所得出的數值3.5±0.6*107M-1相近,證明此種進樣和分離模式可保持生化物種的活性,可用於免疫分析的範疇。在線上混合方面是以梯度方式將10-4M和10-5M Rhodamin B進行混合,所得到的R2值趨近於1,且每一數據點之相對標準偏差小於2%,證明可用於線上反應的定量偵測◦在不連續樣品的分析方面,利用微幫浦推動的動相將微注射器內的樣品帶入至晶片,根據所得到的圖譜,波峰高度和遷移時間的相對標準偏差分別小於8%和0.6%,證明此種方法可與許多流動分析器串接◦
In this thesis, a novel electrophoretic microchip which is capable of directly coupling with flow-through analyzers for uninterrupted sampling was developed. The micro-channel was fabricated on the quartz or the glass substrate by wet etching through Micro-Electro-Mechanical Systems (MEMS) technology. Pressure was used to drive the sample flow through the external tube into the inlet of the microchip and the flow was then split into outlet and electrophoretic channels. A gating voltage was applied to the electrophoretic channel to control the sample loading for subsequent separations and inhibit the sample leakage such that the analysis would not be effected by previous samples. The minimum gating voltage required to inhibit the sample leakage depended on the solution buffer and increased with the hydrodynamic flowrate. This sampling method was applied for the study of affinity interaction between bovine serum albumin (BSA) and its antibody anti-BSA. By introducing a series of samples which contain the mixture of BSA and anti-BSA at different ratios, the binding constant of BSA and anti-BSA was determined to be 3.3 * 107 M-1 which was close to the literature value, 3.5±0.6 * 107 M-1. This result has proved that the developed sampling method conserves the binding activity and can be applied for immuno-analysis. A tee was on-line connected to the sampling line of the microchip and a linear gradient was performed to mix two samples of Rhodamin B at the concentration of 10-4 M and 10-5 M, respectively. The calibration curve constructed based on the results of the gradient was found to have a R2 value close to 1 and the relative standard deviation of each data point was less than 2%, which proved that the sampling method can be used for on-line mixing and quantitative analysis. For the discrete sampling via an on-line injection valve, the relative standard deviation of migration time and peak intensity was determined to be below 0.6% and 8%, respectively. Therefore, such an electrophoresis-based microchip can be directly coupled with any pressure-driven flow analyzers without hardware modifications. To our best knowledge, this is something currently impossible for reported electrophoretic microchip designs.
英文摘要
中文摘要
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一、 陳淑慧,林佾鴻,李國賓,廖寶琦,宋旺洲,林群哲,戴德禮,王蘭玉,蔡銘峰,實驗室晶片“ Lab-On-a-Chip”在生化分析上的應用,中國化工會誌,2001年,第48卷,第三期,39-47.
二、 Yi-Hung Lin,Gwo-Bin Lee,Chun-Wei Li,Guan-Ruey Huang,Shu-Hui Chen,Flow-through Sampling for Electrophoresis-based Microfluidic Chips Using Hydrodynamic Pumping,Journal of Chromatography A ,2001
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