Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/2209
標題: 利用摺疊型微流晶片提升蕪菁黃色嵌紋病毒(TYMV)於感測平面之接附密度與覆蓋均勻度
Improving adhesion density and coverage uniformity of Turnip yellow mosaic virus (TYMV) on the sensor's surface using folded microfluidic channels
作者: 曾秉國
Tseng, Ping-Kuo
關鍵字: T-shaped;摺疊型微流道;U-shaped;W-shaped;microfluidic channel;dipping method;MUA;NCD4;TYMV;浸泡法;MUA;NCD4;TYMV
出版社: 機械工程學系所
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摘要: 
傳統的微生物分子檢測中,其抗原與抗體的接附方法均採用浸泡法。由於布朗運動與擴散效應,微生物分子於接附時會有反應不均勻的現象,本研究利用摺疊型微流晶片來改善上述缺點。
首先藉由有限元素模擬軟體(COMSOL Multiphysics)分析所設計之微流道,並且計算每個區域之流速與流線。利用螢光粒子動態實驗來瞭解微小分子在微流場內移動的軌跡。接著利用傳統浸泡法和摺疊型微流道來接附MUA分子和NCD4螢光於感測晶片表面,並利用共軛焦螢光顯微鏡來量測接附效果。最後以NCD4螢光接附之結果為依據,實際接附TYMV病毒來量測接附效果。
由NCD4螢光接附實驗結果得知,以傳統浸泡法之實驗數據為基準,本論文所設計之摺疊型微流道,平均螢光強度之相對倍率最大為2.67倍;平均螢光覆蓋率之相對倍率最大為2.55倍;螢光覆蓋均勻度最大為80-90%。由TYMV病毒接附實驗結果得知,以傳統浸泡法之實驗數據為基準,本論文所設計之摺疊型微流道,平均螢光強度之相對倍率為3.59倍;平均螢光覆蓋率之相對倍率為19.13倍;螢光覆蓋均勻度為70-90%。相對於傳統浸泡法,摺疊型微流道可以大幅提升MUA、NCD4與TYMV病毒在感測表面的接附密度和覆蓋均勻度。

Recently, there has been an increasing interest to develop rapid, reliable and low-concentration detection methods of microorganisms involved in bioterrorism, food poisoning, and clinical problems. How to detect virus at concentration below the threshold will be challenging with respect to specificity, selectivity, and sensitivity. Among all parameters, sensitivity is probably the most critical consideration. If the sensitivity is not satisfied for real-time detection, researchers need to duplicate numerous numbers of viruses. However, it will substantially increase processing times and experimental hazard. To increase the sensitivity of virus sensors, this paper discusses how to improve adhesion density and coverage uniformity of linkers and virus on the sensor's surface using T-shaped, U-shaped and W-shaped microfluidic channels. In the future, researcher could use emerging technology, such as PT-PCR, QCM, C-V and I-V measurements, etc, to detect viruses on sensor's surface.
Usually microorganisms, molecules, or viruses in the fluidic environment are at very low Reynolds numbers because of tiny diameters. At very low Reynolds numbers, viscous forces of molecules and viruses will dominate. Those micro- or nanoparticles will stop moving immediately when flows cease and drag forces disappear, those phenomena were discovered by the fluorescent particle experiment. Of course, molecules and viruses are still subject to Brownian motion and move randomly. In order to increase the adhesion density of micro- and nanoparticles on sensor's surface, designs of the flow movements in microfluidic channel is proposed.
Adhesion density of linker 11-mercaptoundecanoic acid (MUA) and Turnip yellow mosaic virus (TYMV) with specific quantum dots were measured by confocal microscope. Fluorescent intensity and coverage of quantum dots are used to identify the adhesion density quantitatively. Results show that TYMV and MUA layers disperse randomly by dipping method. Fluorescent intensity of quantum dots; i.e. relative to the amount of MUA and TYMV; were 2.67A.U. and 19.13A.U., respectively, in W-shaped microfluidic devices to contrast just 1.00A.U. and 1.00A.U., respectively, by dipping method. Coverage of MUA and TYMV were 80~90% and 70~90%, respectively, in W-shaped microfluidic channel to contrast just 20~50% and 0~10%, respectively, by dipping method.
URI: http://hdl.handle.net/11455/2209
其他識別: U0005-0608200911180200
Appears in Collections:機械工程學系所

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