簡易檢索 / 詳目顯示

研究生: 蘇琮堯
Su, Tsung-Yao
論文名稱: 以共平面式多電極陣列介電濕潤晶片合成DNA磁球微粒檢測探針之應用
A Multiarray Coplanar-Type-Electrodes EWOD Chip Apply to Magnetic-bead Probes Formation of DNA Detection
指導教授: 饒達仁
Yao, Da-Jeng
口試委員:
學位類別: 碩士
Master
系所名稱: 工學院 - 奈米工程與微系統研究所
Institute of NanoEngineering and MicroSystems
論文出版年: 2010
畢業學年度: 99
語文別: 中文
論文頁數: 119
中文關鍵詞: 介電濕潤生物探針磁球標示
外文關鍵詞: electrowetting on dielectric, EWOD, bio-probe, magnetic-bead labeling
相關次數: 點閱:3下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • 本研究就主要以介電濕潤(Electrowetting-on-dielectric, EWOD)法作為操控微液滴之平台,藉由共平面式多電極之操控陣列可達到同步處理多個DNA檢體,以油包水之操控方式操控包括不同生醫檢體包含:去氧核醣核酸(DNA)、磁球微粒(Magnetic beads)及緩衝液(Buffer)等,並以此完成一可檢測如DNA單點突變(Single-nucleotide Polymorphism, SNP)之磁球探針。
    操控晶片利用ITO(Indium Tin Oxide)玻璃製作出共平面式操控電極,包含多個儲存槽及傳輸電極,搭配無電極之疏水上蓋達到液滴於多個儲存槽同步產生、移動、混合與分離等效果,由於ITO玻璃本身為透明且具有良好之光學性質,因此在後續的光學檢測及影像擷取皆非常便利。透過此共平面式介電濕潤平台的建構,本實驗完成一個開放式的數位微流體系統(Digital Micro Fluidic System),使用者可以更有彈性的進行液珠的傳輸及混合等操控。
    本研究將SNP檢測探針會使用之核酸接合流程在晶片上實行,並對含磁球之液滴做操控測試,最後成功於晶片上完成核酸接合檢測及磁球探針合成步驟,利用量子點(QDots)與探針接合可於螢光顯微鏡下確認探針合成成效,達到實驗室晶片操作目標。


    This study bases on the technique, “Electrowetting on Dielectric (EWOD),” which is actuated by addressable coplanar electrodes for micro droplets manipulation. Multiple bio-reagents such as DNA, magnetic beads and buffer have manipulated synchronous with different concentration. All of the droplets are driven with water oil core-shell type, which are surrounded by silicon oil. All the droplets are generated homogeneously with ultra microvolume(0.3μL).
    A coplanar EWOD chip with several reservoirs and electrodes, is fabricated by indium tin oxide (ITO) glass, which can drive droplets for creating, transporting, merge and splitting. All the fluidics movement is confined between two plates, a bottom plate for the coplanar chip, and another hydrophobic plate with SU8 structure for the top. Detections and capturing of images are easy and convenient through transparent ITO glasses, which are well characteristic of optic. Through the construction of this addressable coplanar-electrode EWOD chip, the study demonstrates a flexible platform for Digital Micro Fluidic (DMF) Systems to manipulate micro droplets.
    To perform a bio-application on the EWOD chip, we transfer a DNA ligation process on this device, which is a common procedure for biomedical analysis. Droplets with magnetic beads are also manipulated on the chip. Moreover, a probe formation process of SNP detection is also completed which confirmed by QDots labeling. The result shows that probes synthesized by EWOD chip are better than by hand. Thus, in this paper, we wish the device could be integrated with other systems to achieve the goal of Micro Total Analysis System (μTAS) and Lab on a Chip.

    摘要 I 目錄 III 圖目錄 VI 表目錄 XIV 第一章 緒論 1 1.1 前言 1 1.2 研究目的 2 第二章 文獻回顧 4 2.1 介電濕潤現象與微液滴之操控 4 2.2 微液滴之混合與介電濕潤之探討 10 2.3 不同介電材料對介電濕潤之探討 15 2.4 介電濕潤系統於生醫操作上之應用 23 2.5 磁球與酵素之生物感測 41 2.6 文獻總結 41 第三章 研究方法 43 3.1 實驗原理 43 3.1.1 介電濕潤原理 43 3.1.2 介電濕潤晶片製程 44 3.1.3 介電層材料選擇 51 3.1.4 實驗系統架設 54 3.2 液滴操控與生醫檢體配製設計 55 3.2.1 共平面式晶片液滴操控 55 3.2.2 磁球液滴操控與收集 56 3.2.3 DNA Ligation與SNP磁球探針合成設計 58 第四章 實驗成果 66 4.1 共平面式介電濕潤晶片與疏水上蓋製程結果 66 4.1.1 共平面式晶片與疏水上蓋製作 66 4.1.2 接觸角量測 68 4.2 微液滴之基礎操控 69 4.2.1 DI水操控測試 69 4.3 磁球液滴與核酸接合實驗 78 4.3.1 磁球液滴操控實驗 78 4.3.2 核酸接合(DNA ligation)實驗 83 4.4 磁球探針合成實驗 89 4.4.1 緩衝溶液(Wash Bfr. & Binding Bfr.)操控測試 90 4.4.2 SOG(Spin on glass)之應用與磁球探針合成 93 4.5 磁球探針合成確認與量子點螢光觀測 103 第五章 結論與未來展望 115 5.1 結論 115 5.2 未來展望 116 第六章 參考文獻 117

    [1] R. Feynman, "There's plenty of room at the bottom," Journal of Microelectromechanical Systems, vol. 1, pp. 60-66, Mar. 1992.
    [2] R. Feynman, "Infinitesimal machinery," Journal of Micro Electro Mechanical Systems, vol. 2, pp. 4-14, Mar. 1993.
    [3] D. R. Reyes, D. Iossifidis, P.-A. Auroux, and A. Manz, "Micro Total Analysis Systems. 1. Introduction, Theory, and Technology," Analytical Chemistry, vol. 74, pp. 2623-2636, 2002.
    [4] P. A. Auroux, D. Iossifidis, D. R. Reyes, and A. Manz, "Micro Total Analysis Systems. 2. Analytical Standard Operations and Applications," Analytical Chemistry, vol. 74, pp. 2637-2652, 2002.
    [5] S. J. Lee and S. Y. Lee, "Micro total analysis system (µ-TAS) in biotechnology," Applied Microbiology and Biotechnology, vol. 64, pp. 289-299, 2004.
    [6] S. K. Cho, H. Moon, J. Fowler, and C. J. Kim, "Splitting a liquid droplet for electrowetting-based microfluidics," in ASME International Mechanical Engineering Congress and Exposition. vol. IMECE2001/MEMS-23830 New York, 2001.
    [7] M. G. Pollack, R. B. Fair, and A. D. Shenderov, "Electrowetting-based actuation of liquid droplets for microfluidic applications," Applied Physics Letters, vol. 77, pp. 1725-1726, 2000.
    [8] S. K. Cho, M. Hyejin, and C. J. Kim, "Creating, transporting, cutting, and merging liquid droplets by electrowetting-based actuation for digital microfluidic circuits," Microelectromechanical Systems, Journal of, vol. 12, pp. 70-80, 2003.
    [9] J. Fowler, M. Hyejin, and K. Chang-Jin, "Enhancement of mixing by droplet-based microfluidics," in Micro Electro Mechanical Systems, 2002. The Fifteenth IEEE International Conference on, 2002, pp. 97-100.
    [10] P. Paik, V. K. Pamula, M. G. Pollack, and R. B. Fair, "Electrowetting-based droplet mixers for microfluidic systems," Lab on a Chip, vol. 3, pp. 28-33, 2003.
    [11] C. G. Cooney, C. Y. Chen, M. R. Emerling, A. Nadim, and J. D. Sterling, "Electrowetting droplet microfluidics on a single planar surface," Microfluid Nanofluid, vol. 2, pp. 435-446, 2006.
    [12] J. Berthier, Microdrops and Digital Microfluidics. NY, USA: William Andrew Inc., 2008.
    [13] A. G. Papathanasiou, A. T. Papaioannou, and A. G. Boudouvis, "Illuminating the connection between contact angle saturation and dielectric breakdown in electrowetting through leakage current measurements," Journal of Applied Physics, vol. 103, p. 034901, 2008.
    [14] S. K. Fan, H. Yang, T. T. Wang, and W. Hsu, "Asymmetric electrowetting-moving droplets by a square wave," Lab on a Chip, vol. 7, pp. 1330-1335, 2007.
    [15] L. Yifan, W. Parkes, L. I. Haworth, A. Ross, J. Stevenson, and A. J. Walton, "Room-Temperature Fabrication of Anodic Tantalum Pentoxide for Low-Voltage Electrowetting on Dielectric (EWOD)," Microelectromechanical Systems, Journal of, vol. 17, pp. 1481-1488, 2008.
    [16] L. Yifan, Y. Mita, L. I. Haworth, W. Parkes, M. Kubota, and A. J. Walton, "Test Structure for Characterizing Low Voltage Coplanar EWOD System," Semiconductor Manufacturing, IEEE Transactions on, vol. 22, pp. 88-95, 2009.
    [17] V. Srinivasan, V. K. Pamula, M. G. Pollack, and R. B. Fair, "Clnical Diagnostics on Human Whole Blood, Plasma, Serum, Urine, Saliva, Sweat, and Tears on A Digital Microlfuidic Platform."
    [18] L. Lei, H. Huan, L. Hong, and Y. Da-tian, "Electrowetting of the blood droplet on the hydrophobic film of the EWOD chips," in Engineering in Medicine and Biology Society, 2005. IEEE-EMBS 2005. 27th Annual International Conference of the, 2005, pp. 1941-1944.
    [19] D. Brassard, L. Malic, F. Normandin, M. Tabrizian, and T. Veres, "Water-oil core-shell droplets for electrowetting-based digital microfluidic devices," Lab on a Chip, vol. 8, pp. 1342-1349, 2008.
    [20] L. Malic, T. Veres, and M. Tabrizian, "Biochip functionalization using electrowetting-on-dielectric digital microfluidics for surface plasmon resonance imaging detection of DNA hybridization," Biosensors and Bioelectronics, vol. 24, pp. 2218-2224, 2009.
    [21] R. Sista, Z. Hua, P. Thwar, A. Sudarsan, V. Srinivasan, A. Eckhardt, M. Pollack, and V. Pamula, "Development of a digital microfluidic platform for point of care testing," Lab on a Chip, vol. 8, pp. 2091-2104, 2008.
    [22] G. J. Shah and C. J. Kim, "Meniscus-Assisted High-Efficiency Magnetic Collection and Separation for EWOD Droplet Microfluidics," Microelectromechanical Systems, Journal of, vol. 18, pp. 363-375, 2009.
    [23] Y. J. Liu, D. J. Yao, H. C. Lin, W. Y. Chang, and H. Y. Chang, "DNA ligation of ultramicro volume using an EWOD microfluidic system with coplanar electrodes," Journal of Micromechanics and Microengineering, vol. 18, pp. 1-7, Apr 2008.
    [24] H. C. Lin, Y. J. Liu, D. J. Yao, and H. Y. Chang, "Ewod based microfluidic system with coplanar electrodes for parallel DNA ligation of ultramicro volume," in Solid-State Sensors, Actuators and Microsystems Conference, 2009. TRANSDUCERS 2009. International, 2009, pp. 92-95.
    [25] H.C. Lin, Y.J. Liu, and D.J. Yao, "Core-Shell Droplets for Parallel DNA Ligation of an Ultra-micro Volume Using an EWOD Microfluidic System," Journal of the Association for Laboratory Automation, vol. 15, pp. 210-215, 2010.
    [26] Y. S. Huang, Y. H. Peng, and S. K. Fan, "Microfluidic immunosensor based on insulator dielectrophoresis and electrowetting-on-dielectric," in Nano/Micro Engineered and Molecular Systems (NEMS), 2010 5th IEEE International Conference on, 2010, pp. 337-339.
    [27] M. H. Liao, J. C. Guo, and W. C. Chen, "A disposable amperometric ethanol biosensor based on screen-printed carbon electrodes mediated with ferricyanide-magnetic nanoparticle mixture," Journal of Magnetism and Magnetic Materials, vol. 304, pp. 421-423, 2006.
    [28] B. W. Lu and W. C. Chen, "A disposable glucose biosensor based on drop-coating of screen-printed carbon electrodes with magnetic nanoparticles," Journal of Magnetism and Magnetic Materials, vol. 304, pp. 400-402, 2006.
    [29] Z. Liu, Y. Liu, H. Yang, Y. Yang, G. Shen, and R. Yu, "A phenol biosensor based on immobilizing tyrosinase to modified core-shell magnetic nanoparticles supported at a carbon paste electrode," Analytica Chimica Acta, vol. 533, pp. 3-9, 2005.
    [30] Y. J. Liu, "DNA Reaction and Detection using an EWOD Microfluidic System with Coplanar Electrodes," in Nano Engineering and Microsystems. vol. Doctor Hsinchu, Taiwan: National Tsing Hua University, 2009, p. 79.
    [31] N. N. Toan, "Spin-on glass materials and applications in advanced IC technologies," Hanoi, Vietnam, 1999.

    無法下載圖示 全文公開日期 本全文未授權公開 (校內網路)
    全文公開日期 本全文未授權公開 (校外網路)

    QR CODE