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Title: | 利用螢光手機顯微鏡結合濃縮晶片量測粒子的擴散行為並用於C反應蛋白檢測 Measurement of Nano-particle’s Diffusion Behaviors for Detecting Concentration of CRP in Preconcentration Chip by Fluorescent Cellphone Microscope |
Authors: | Yi-Fan Fang 方依凡 |
Advisor: | 沈弘俊(Horn-Jiunn Sheen) |
Keyword: | 手機顯微鏡,布朗運動,微流道閥門,預濃縮晶片,螢光相關光譜,C反應蛋白,生物感測技術, Cellphone Microscope,Brownian Motion,Micro-channel Valve,Preconcentration Chip,Fluorescence Correlation Spectroscopy,C-reactive protein,Biosensor, |
Publication Year : | 2017 |
Degree: | 碩士 |
Abstract: | 本研究成功開發了一種新型可攜式生醫裝置及檢測方法,包括手機顯微鏡(Cellphone Microscope)、螢光相關光譜(Fluorescence Correlation Spectroscopy)、預濃縮晶片(Preconcentration Chip)三大部分,利用此裝置,偵測不同濃度之C反應蛋白 (C Reactive Protein, CRP)接於修飾有CRP抗體(anti-CRP)的螢光粒子時,會使粒子粒徑變大,導致其布朗運動擴散係數(Diffusion Coefficient)下降,再結合奈米預濃縮晶片,可以大幅提升此裝置檢測極限。
本研究分為三大部分:螢光手機顯微鏡部分,使用華碩 Zenfon Zoom作為影像擷取裝置,透過光路設計及利用凹、凸透鏡組結合成放大鏡頭(External lens),可以大幅降低像差、色差,搭配一般藍紫光雷射筆(405 nm)作為激發光源,可構成一光學放大倍率約為5X的螢光顯微鏡系統;螢光相關光譜部分,計算區域內螢光亮度消散之自相關性,再以曲線擬合求得特徵擴散時間,進而求得區域內粒子之擴散係數,作為布朗運動定量分析的重要指標;預濃縮晶片部分,利用微影製程,製作出微米級流道,再以多孔性奈米材料Nafion作為奈米流道、離子選擇薄膜(ion-selective membrane),其中主流道上設計有一長寬皆為600 µm之艙體,此為檢測區塊,而區塊之兩旁,又設計兩個細小狹縫,連接同一個進口,而此進口通入PDMS-A劑,作為流體閥門流道,用於捕捉檢測區塊。 實驗時,施加電位差於Nafion的兩端使產生濃度極化效應(ion concentration polarization, ICP),此時一端為離子富集區(ion enrichment region)另一端為離子空乏區(ion depletion region),接者在空乏區一側施加偏壓,便可利用第二種電滲流 (electroosmosis of the second kind)使濃縮區塊(preconcentration plug)產生,再利用流道的特殊設計,使濃縮區塊能停滯在檢測區塊中,並利用閥門捕捉,進行螢光手機顯微鏡拍攝15分鐘,進而分析其擴散係數變化。 此研究成功提供了一種快速、廉價、及時的蛋白質或病毒感測方法,且再經過濃縮晶片濃縮機制,使極低濃度之CRP (pg/ml)亦可檢測,大幅提升了檢測極限。 In this study, we successfully develop a new type of portable biosensor, including Cellphone Microscope, Fluorescence Correlation Spectroscopy and Preconcentration Chip three parts. We can use this device to detect different concentrations of CRP by connecting CRP antibody to fluorescent particles, due to the particle size will become larger, it will result in its Brownian motion diffusion coefficient decreased. This device will enhance the detection limitation greatly by combining with Preconcentrtion chip. This study is divided into three parts: (1) Fluorescent Cellphone Microscope, we use ASUS Zenfone Zoom as CCD and reduce the aberration significantly by using concave and convex lens group as external lens. Then, we choose 405nm laser pointer as the excitation light source. It can form a Fluorescenct Cellphone Microscope system with an optical magnification about 5X. (2) Fluorescence Correlation Spectroscopy, it calculates the autocorrelation of the particles’ fluorescence intensity dissipation in the region, and then obtain the characteristic diffusion time to get the diffusion coefficient. (3) Preconcentration chip, it is fabricated through the lithography process. We use Nafion as ion-selective membrane and then couple with the uncured PDMS fluid valve to capture the enrichment detection block. In this experiment, we apply voltage drop to generate ion concentration polarization(ICP) effect. At this point, the one side of Nafion is ion enrichment region, and the other is ion depletion region. At this moment, we apply voltage drop in the depletion side, and the preconcentration plug will be produced by using electroosmosis of the second kind. And then, we catch the plug by the uncured PDMS fluid valve to detect the change of the Brownian motion diffusion coefficient using Fluorescent Cellphone Microscope. In conclusions, we developed an fast, cheap and real-time way to detect proteins or virus and then through the concentration enrichment mechanism, so that very low concentration of CRP (pg / ml) can also be detected. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77946 |
DOI: | 10.6342/NTU201702117 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 應用力學研究所 |
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