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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/49846

Title: 微流道生物晶片之核苷酸雜合綜合分析
A Comprehensive Analysis of Microfluidic Biochips through DNA Hybridization
Authors: Lin, Chia-Ping
Contributors: NTOU:Department of Electrical Engineering
Keywords: DNA雜合;雷射激發螢光;PMMA(聚甲基丙烯酸甲酯);微流道生物晶片;倒立式螢光顯微鏡
DNA hybridization;laser-induced fluorescence;PMMA (Poly methyl methacrylate);microfluidics biochips;inverted fluorescence microscope
Date: 2017
Issue Date: 2018-08-22T07:13:06Z
Abstract: 本論文利用PMMA(Poly methyl methacrylate)聚甲基丙烯酸甲酯之高分子聚合物作為實驗研究用晶片基材,透過微機電製程技術製作出簡易型核苷酸雜合微流道之生物晶片。 在晶片製作上,利用CO2雷射雕刻機在基材上雕刻出生物晶片之微流道結構,加工後的基材再經過中性清潔劑、去離子水以及異丙酮清洗、烘烤、UV紫外光照射表面改質、熱壓成一體完工之實驗用晶片。 生物晶片最主要結構為第三層的主腔室,以帶有螢光探針之DNA注入晶片裡,利用倒立式螢光顯微鏡觀察腔室周圍DAN分子移動變化,注入界面活性劑通過晶片內部DNA樣品利用螢光變化,藉以區别判斷樣品雜合和不雜合。另外再使用另台螢光檢測系統檢測,架構為波長532nm綠光雷射以及550nm的高通濾光片與PSoC,即時迅速的反應出樣品的電壓訊號。 在實驗的樣品上,採用不同的微生物菌種培養,以親水性產氧單胞菌、愛德華氏菌、克雷伯氏肺炎菌、大腸桿菌等,將樣品透過微流道轉入晶片裡的主腔室,再利用螢光探針針對不同微生物樣品的專一性,在倒立式螢光顯微鏡下固定以最佳濃度百分比界面活性劑沖刷進行雜合判斷。
This thesis conducts an experimental research on microfluidic biochips. The experiments use macromolecular polymers of PMMA (Poly methyl methacrylate) as the substrate of biochips and apply Micro-Electromechanical Systems (MEMS) to produce handy microfluidic biochips capable of performing DNA hybridization. With the help of CO2 laser engraving machines, we engrave the microfluidic structure of biochips on the substrate. Then we wash the engraved substrate with neutral detergent, deionized water, and IPA before applying it to be heated, UV-Ozone cleaned, and bonded to turn into chips suitable for our experiments. A place in the primary part of a biochip, or the cavity chamber. Then we instill the chemical agent TE3 into the activated DNA with Probe-HEX into the biochip. We can see clearly a ring of bright DNA around and using an inverted fluorescence microscope. Further, we instill surfactants into primary part of a biochip, which causes DNA molecules with different characteristics to move at different speeds. In this way, we may tell whether the sample is hybridized or not. In addition, we use the fluorescent system with the setting of 532nm wavelength with Green Laser, 550nm high-pass filter, and PSoC, to measure the piezoelectric signal of the sample immediately. We incubate different microorganism cultures, such as Aeromonas hydrophila, Edwardsiella tarda, Klebsiella pneumoniae, and Escherichia coli, to serve as the samples blotted in the biochips. Given the specific reactions of Probe-HEX to different microorganistic samples, we use surfactants of the most applicable percentages of concentration to cleanse the samples and diagnose their hybridization.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010353065.id
Appears in Collections:[電機工程學系] 博碩士論文

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