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研究生: 陳信彬
Shin-Bin Chen
論文名稱: 開發一適用於細胞分選之掃流式過濾晶片
The Development of a Filtration Chip Based on Crossflow Filtration for Cell Separation and Collection
指導教授: 陳品銓
Pin-Chuan Chen
口試委員: 陳品銓
Pin-Chuan Chen
張復瑜
Fuh-Yu Chang
王孟菊
Meng-Jiy Wang
饒達仁
Da-Jeng Yao
曹嘉文
Chia-Wen Tsao
學位類別: 碩士
Master
系所名稱: 工程學院 - 機械工程系
Department of Mechanical Engineering
論文出版年: 2019
畢業學年度: 107
語文別: 中文
論文頁數: 118
中文關鍵詞: 微型過濾晶片掃流式過濾細胞分選黏合
外文關鍵詞: microfiltration chip, Cross-flow filtration, Cell sorting, Bonding
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  • 本研究為開發一微型過濾晶片,利用掃流式過濾原理,可用於三種不同尺寸範圍的粒子分離,達成細胞分選之目的,使用商用濾膜將其整合於晶片中,此濾膜擁有成本低廉、易於取得的優點,然而此種濾膜為核孔聚碳酸酯(Nucleopore Polycarbonate Track-Etch)與晶片基材聚甲基丙烯酸甲酯 (PMMA, Polymethyl methacrylate)不同,在晶片封裝時屬於異質黏合,製程較為複雜,故本研究提出(1)以UV光固化膠黏合製程進行黏合兩種不同的材質,並透過螺旋型流道幾何改善檢體於晶片內流動情況,以降低檢體流動不順的現象;(2)雙層掃流式的結構設計,可使粒子不易阻塞於濾膜前,改善過濾時壓力過大等問題。為了提高過濾晶片的過濾效率,實驗將針對不同尺寸的流道設計進行測試,經由計算過濾效率,並分析其影響;再以人類全血作為分析濾膜孔徑對於血細胞的分離效率之影響,以及測量細胞的存活率。實驗結果顯示,流道寬度2 mm、流道深度3 mm之晶片,濾膜孔徑尺寸為12 um、8 um,三種尺寸範圍的粒子分離效率分別為96.9、33.27、78.23%。在血液測試的實驗結果顯示,流道寬度2 mm、流道深度3 mm之晶片,濾膜孔徑尺寸為5 um、0.8 um,可成功攔截67.83%的白血球、72.83%的紅血球、以及蒐集60%的血漿;細胞存活率皆維持在約90%。


    The main purpose of this study is to develop a microfluidic chip, which can separate three different sizes of particles by the principle of cross-flow filtration, and achieve the purpose of cell separating. The filter integrated in the chip is a commercial filter, and the cost is low. The advantages are easy to obtain. However, the material of the filter is different from the chip substrate. It is heterogeneously bonded during chip packaging, and the process is complicated. In this study, two different materials are bonded by UV glue bonding process. The spiral channel improves the flow of the sample in the chip and reduces the phenomenon that the sample flow is not smooth. The double cross-flow filtration structure can make the particles not easily block on the filter, and improve the problems caused by excessive pressure. In order to improve the filtration efficiency of the filter chip, the experiment will test the microfluidic chip design of different sizes, calculate the filtration efficiency, and analyze its influence. Finally, the human’s whole blood is used as the sample to analyze the effect the membrane pore size on the separation efficiency of blood cells. And measure the survival rate of the cells. In the results 2 mm flow path width, 3 mm flow path depth of the wafer, the size of the filter is 12 um and 8 um, the three different sizes of particle separation efficiency are 96.9%, 33.27% and 78.23%. In the blood test experiment 2 mm flow path width, 3 mm flow path depth of the wafer, the size of filter is 5 um and 0.8 um, can intercept 67.83% of white blood cells, 72.83% of red blood cells, and collect 60% of plasma; cell viability was maintained at around 90%.

    目錄 摘要 I Abstract III 誌謝 V 目錄 VII 圖目錄 XI 表目錄 XV 符號表 XVI 第一章 導論 1 1.1研究背景 1 1.2研究動機 2 1.3研究方法 5 1.4論文架構 6 第二章 文獻回顧 9 2.1主動式分離技術 9 2.2被動式分離技術 11 2.3掃流式過濾 16 第三章 過濾晶片初步設計及結果 21 3.1晶片設計介紹 21 3.2結構設計 22 3.3流動情況相關設計 25 第四章 晶片製程介紹 27 4.1 微銑削 27 4.1.1前言 27 4.1.2 操作與使用方法 29 4.2晶片製造 32 4.2.1上層上蓋、下蓋晶片加工 34 4.2.2上層下蓋背部加工 36 4.2.3下層上蓋、下蓋晶片加工 37 4.2.4出口流道晶片加工 39 4.3晶片黏合 40 4.3.1塑膠材料化學黏合 40 4.3.2 UV膠黏合 43 第五章 研究設備與實驗方法 45 5.1研究設備 45 5.1.1製程設備與軟體 45 5.1.2量測設備與軟體 49 5.2實驗方法 54 5.2.1.1流速對過濾能力之影響 55 5.2.1.2影像處理 57 5.2.2流道深寬比對過濾效率之影響 60 5.2.3流道寬對過濾效率之影響 62 5.2.4血液分離實驗 64 5.2.5血細胞存活率測試 67 第六章 實驗結果與討論 69 6.1流速對過濾能力之影響結果 69 6.2流道深寬比例對過濾效率影響之結果 73 6.3流道寬對過濾效率影響之結果 75 6.4血液分離實驗結果 77 6.5血細胞存活率測試結果 81 第七章 結論與未來展望 83 7.1結論 83 7.2未來展望 86 參考文獻 89 附錄A 流速對過濾能力之影響實驗數據 97 附錄B 流道深寬比對過濾效率之影響實驗數據 98 附錄C流道寬對過濾效率之影響實驗結果 101 附錄D血液分析數據(12 µm、0.8 µm) 102 附錄E血液分析數據(8 µm、0.8 µm) 103 附錄F血液分析數據(5 µm、0.8 µm) 105

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