Responsive image
博碩士論文 etd-0723109-084810 詳細資訊
Title page for etd-0723109-084810
論文名稱
Title
溫度變化對DNA分子於漸縮/及漸擴微流道之拉伸及形變研究
Stretching and Deformation of DNA Molecules in a Converging-Diverging Microchannel with Heating Effect
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
97
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2009-07-02
繳交日期
Date of Submission
2009-07-23
關鍵字
Keywords
水力拉伸、熱效應、DNA分子、共軛焦雷射掃描顯微鏡、微質點影像測速儀
thermal effects, μPIV, conformation and dynamics, DNA molecule, CLSM
統計
Statistics
本論文已被瀏覽 5722 次,被下載 1888
The thesis/dissertation has been browsed 5722 times, has been downloaded 1888 times.
中文摘要
本實驗設計適合加熱且觀測之漸縮漸擴微流道,其縮擴比為8:1,以電動驅動DNA溶液,控制電場強度小於150 V/cm,以避免產生焦耳熱效應,期望產生一均勻延展流及齊次的應變率,並在微流道上方利用等熱通量的方式對微流道內部之緩衝溶液進行加熱,搭配熱電隅及雷射誘發微螢光(Micro Laser-Induced Fluorescence)量測液體溫度,控制緩衝溶液溫度為25、35、45及55°C,避免過高的溫度使DNA分子受損。利用微質點影像測速儀(Micrometer resolution Particle Image Velocimetry) 計算局部應變率,評估DNA伸長量,再配合螢光可視化的量測技術,搭配共軛焦雷射掃瞄顯微鏡(Confocal Laser Scanning Microscopy) 進行拍攝,在不同的緩衝溶液溫度下,統計DNA在微流道中拉伸長度之分佈,並計算DNA分子承受之流體阻力,及其所造成的形態變化和應力鬆弛時間,希望藉此對溫度影響DNA分子於微流道中拉伸及形變有概括的了解。
Abstract
In this study, an electrokinetics-induced elongation flow was created inside a gradual converging-diverging microchannel with different temperature (25, 35, 45, 55°C). The conformation of DNA molecules, local strain rate, and the relaxation time play important roles in determining the extent of DNA stretching. By using μPIV/μLIF measurements, the velocity/temperature distributions in microchannels can be secured. The local strain rate was estimated by μPIV measurements. We observe the hydrodynamic stretching DNA molecules in elongation flow by confocal laser scanning microscope (CLSM). Through CLSM images analysis, relaxation time of DNA molecules was estimated. Finally, dynamic properties and stretching ratio of DNA molecules stretched by EOF driven at various electric field and temperature ware measured. The thermal effect and the electric field on the conformation were also studied and discussed.
目次 Table of Contents
目錄 i
表目錄 iii
圖目錄 iv
符號說明 vi
中文摘要 viii
Abstract ix
第一章 序論 1
1-1 前言 1
1-2 DNA分子結構與性質 1
1-3 生醫晶片 2
1-4 背景與目的 3
1-5 文獻回顧 5
第二章 實驗系統與設備 12
2-1 μPIV及μLIF系統 12
2-2 共軛焦雷射掃瞄顯微鏡量測系統 13
2-3 直流電源供應器 14
2-4 製程設備 14
2-5 其他實驗週邊設備 16
第三章 實驗方法及步驟 26
3-1 微流道設計與製程 26
3-2 工作流體配製 29
3-3 加熱系統建立 29
3-4 光學量測系統建立及原理 30
3-5 實驗量測參數 33
第四章 理論分析 43
4-1 無因次參數分析 43
4-2 DNA分子水力拉伸形變 45
第五章 誤差分析 48
第六章 結果與討論 52
6-1 μPIV之流場量測 52
6-2 μLIF之溫度場量測 52
6-3 DNA螢光可視化觀測 53
6-4 DNA分子受力分析 55
6-5 DNA分子拉伸模型分析 57
第七章 結論與建議 71
7-1 結論 71
7-2 建議與改進 72
參考文獻 74
附錄 A 80
參考文獻 References
1. S. Chu, "Laser Manipulation of Atoms and Particles," Science, Vol. 253, 1991, pp. 861-866.
2. T. T. Perkins, D. Smith, and S. Chu, "Direct Observation of Tube-like Motion of a Single Polymer Chain," Science, Vol. 264, 1994, pp. 819-822.
3. T. T. Perkins, S. Quake, D. Smith, and S. Chu, "Relaxation of a Single DNA Molecule Observed by Optical Microscopy," Science, Vol. 264, 1994, pp. 822-826.
4. C. Bustamante, J. F. Marko, E. D. Sigia, and S. Smith, "Entropic Elasticity of lambda-phage DNA," Science, Vol. 265, 1994, pp. 1599-1600.
5. T. T. Perkins, D. E. Smith, R. G. Larson, and S. Chu, "Stretching of a Single Tethered Polymer in a Uniform Flow," Science, Vol. 268, 1995, pp. 83-87.
6. P. Cluzel, A. Lebrun, C. Heller, R. Lavery, J. L. Viovy, D. Chatenay, and F. Caron, "DNA: an extensible molecule," Science, Vol. 271, 1996, pp. 792-794.
7. T. Strick, J. F. Allemand, D. Bensimon, A. Bensimon, and V. Croquette, "The Elasticity of a Single Supercoiled DNA Molecule," Science, Vol.271, 1996, pp. 1835-1837.
8. C. Gosse, and D. Wirtz, "Magnetic tweezers for DNA Micromanipulation," Rev. Sci. Instrum., Vol. 71, 2000, pp. 4561-4570.
9. S. B. Smith, L. Finzi, and C. Bustamante, "Direct Mechanical Measurements of The Elasticity of Single DNA Molecules by Using Magnetic Beads," Science, Vol. 258, 1992, pp. 1122-1126.
10. C. Gosse, and V. Croquette, "Magnetic tweezers: Micromanipulation and Force Measurement at the Molecular Level," Biophys. J, Vol. 82, 2002, pp. 3314-3329.
11. P. G. De Gennes, "Coil-Stretch Transition of Dilute Flexible Polymers Under Ultrahigh Velocity Gradients," J. Chem. Phys. Vol. 60, 1974, pp. 5030-5042.
12. M. J. Menasveta and D. A. Hoagland, "Light Scattering from Dilute Polystyrene Solutions in Uniaxial Elongational Flow," Macromolecules, Vol. 24, 1991, pp. 3427-3433.
13. D. E. Smith, H. P. Babcock, and S. Chu, "Single-Polymer Dynamics in Steady Shear Flow," Science, Vol. 283, 1999, pp. 1724-1727.
14. E. C. Lee, M, J, Solomon, and S. J. Muller, "Molecular Orientation and Deformation of Polymer Solutions Under Shear: A Flow Light Scattering Study," Macromolecules, Vol. 30, 1997, pp. 7313-7321.
15. E. C. Lee and S. J. Muller, "Flow Light Scattering Studies of Polymer Coil Conformation in Solutions in Extensional Flow," Macromolecules, Vol. 32, 1999, pp. 3295-3305.
16. D. E. Smith and S. Chu, "Response of Flexible Polymers to a Sudden Elongational Flow," Science, Vol. 281, 1998, pp. 1335-1339.
17. J. L. Lumley, "Drag Reduction by Additives," Ann. Rev. Fluid Mech., Vol. 1, 1969, pp. 367-383.
18. P. LeDuc, C. Haber, G. Bao, and D. Wirtz, "Dynamics of Individual Flexible Polymers in a Shear Flow," Nature, Vol. 399, 1999, pp. 564-566.
19. G. H. Mckinley and T. Sridhar, "Filament-Stretching Rheometry of Complex Fluids," Ann. Rev. Fluid Mech., Vol. 34, 2002, pp. 375-415.
20. R. G. Larson, "The Structure and Rheology of Complex Fluid," Oxford University Press, 1999, New York.
21. S. S. Hsieh, C. H. Liu, and J. H. Liou, "DNA Molecules Dynamics in a Cross-Slot Microchannel," Meas. Sci. Technol., Vol. 18, 2007, 2907-2915.
22. G. G. Fuller, "Optical Rheometry of Complex Fluids," Oxford University Press, 1995, New York.
23. H. C. Ottinger, "Stochastic Processes in Polymeric Fluids," Springer, 1996, Berlin.
24. S. B. Smith, Y. Cui, and C. Bustamante, "Overstretching B-DNA: The Elastic Response of Individual Double-Stranded and Single-Stranded DNA Molecules," Science, Vol. 271, 1996, pp.795-799.
25. T. T. Perkins, D. E. Smith, and S. Chu, "Single Polymer Dynamics in an Elongational Flow," Science, Vol. 276, 1997, pp. 2016-2021.
26. D. E. Smith and S. Chu, "Response of Flexible Polymers to a Sudden Elongational Flow," Science, Vol. 281, 1998, pp. 1335-1340.
27. P. S. Doyle and B. Ladoux, "Stretching Tethered DNA Chains in Shear Flow," Europhys. Lett., Vol. 52, 2000, pp. 511-517.
28. Z. Ronai, C. Barta, and A. Guttman, "DNA Analysis on Electrophoretic Microchips: Effect of Operational Variables," Electrophoresis, Vol. 22, 2001, pp. 294-299.
29. M. C. Williams, J. R. Wenner, L. Rouzina, and V. A. Bloomfield, "Entropy and Heat Capacity of DNA melting from Temperature Dependence of Single Molecule Stretching," Biophys. J., Vol. 80, 2001, pp. 1932-1939.
30. G. C. Randall and P. S. Doyle, "Electrophoretic Collision of a DNA Molecule with an Insulating Post," Phys. Rev. Lett., Vol. 93, 2004, pp. 58102-1.
31. J. B. Heng, A. Aksimentiev, C. Ho, P. Marks, Y. V. Grinkova, S. Sligar, K. Schulten, and G. Timp, "Stretching DNA Using the Electric Field in a Synthetic Nanopore," Nano Lett., Vol. 5, 2005, pp. 1883-1888.
32. S. Jeong, S. K. Park, J. K. Chang, and S. H. Kang, "Ultra-Sensitive Real-Time Single-DNA Molecules Detection at a Fused-Silica/Water Interface Using TIRFM Technique," Bull. Korean Chem. Soc., Vol. 26, 2005, pp. 979-982.
33. J. W. Larson, G. R. Yantz, Q. Zhong, R. Charnas, C. M. D’Antoni, M. V. Gallo, K. A. Gillis, L. A. Neely, K. M. Phillips, G. G. Wong, S. R. Gullans, and R. Gilmanshin, "Single DNA Molecule Stretching in Sudden Mixed Shear and Elongational Microflows," Lab Chip, Vol. 6, 2006, pp.1187-1199.
34. B. Yang, V. R. Dukkipati, D. Li, B. L. Cardozo, and S. W. Pang, "Stretching and Selective Immobilization of DNA in SU-8 Micro- and Nanochannels," J. Vac. Sci. Technol., Vol. 25, 2007, pp. 2352–2356.
35. J. Tang and P. S. Doyle, "Electrophoretic Stretching of DNA Molecules Using Microscale T Junctions," Appl. Phys. Lett., Vol. 90, 2007, pp. 224103.
36. M. Ichikawa, H. Ichikawa, K. Yoshikawa, and Y. Kimura, "Extension of a DNA Molecule by Local Heating with a Laser," Phys. Rev. Lett., vol. 99, 2007, pp. 148104.
37. K. Yamashita , M. Miyazaki, Y. Yamaguchi, M. P. Briones , H. Nakamura , and H. Maeda, "Direct Circular Dichroism Spectra Measurement of Stretching Long-strand DNA in a Tapering Microchannel," Chem. Eng. J., Vol.135, 2008, pp. S288–S291.
38. M. C. J. Coolen, R. N. Kieft, C. C. M. Rindt, and A. A. Van Steenhoven, "Application of 2-D LIF Temperature Measurements in Water Using a Nd:YAG Laser," Exp. Fluids, Vol. 27, 1999, pp. 420-426.
39. D. Ross, M. Gaitan, and L. E. Locascio, "Temperature Measurement in Microfluidic Systems Using a Temperature-Dependent Fluorescent Dye, "Analytical Chemistry, Vol. 73, 2001, pp. 4117-4123.
40. 伍秀菁, 汪若文, 林美吟編輯, "微機電系統技術與應用," 國科會精密儀器發展中心出版, 2003.
41. S. S. Hsieh and J. H. Liou, "DNA Molecule Dynamics in Converging-Diverging Microchannels," Biotechnol. Appl. Biochem., Vol. 52, 2009, pp. 29-40.
42. G. C. Randall and P. S. Doyle, "Collision of a DNA Polymer with a Small Obstacle," Macromolecules, Vol. 39, 2006, pp. 7734-7745.
43. P. D. Patel and Eric S. G. Shaqfeha, "A Computational Study of DNA Separations in Sparse Disordered and Periodic Arrays of Posts, " J. Chem. Phys., Vol. 118, 2003, pp. 2941-2951.
44. R. G. Larson, "The Rheology of Dilute Solutions of Flexible Polymers: Progress and Problems," Journal of Rheology, Vol. 49, 2005, pp. 1-70.
45. P. J. Shrewsbury, S. J. Muller, and D. Liepmann, "Effect of Flow on Complex Biological Macromolecules in Microfluidic Devices," Biomedical. Microdevices, Vol. 3, 2001, pp. 225-238.
46. J. R. Taylor, "An Introduction to Error Analysis," University Science Books, Sausalito, 1997.
47. C. C. Zuo, F. Ji, Q. Q. Cao, and J. S Yang, "Numerical Study on Electrophoretical Stretching Dynamics of DNA in Microcontraction," Polymer, Vol. 49, 2008, pp. 809-815.
48. J. H. Kim, W. X. Shi, and R. G. Larson, "Methods of Stretching DNA Molecules Using Flow Fields," Langmuir, Vol. 23, 2007, pp. 755-764.
電子全文 Fulltext
本電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。
論文使用權限 Thesis access permission:校內校外完全公開 unrestricted
開放時間 Available:
校內 Campus: 已公開 available
校外 Off-campus: 已公開 available


紙本論文 Printed copies
紙本論文的公開資訊在102學年度以後相對較為完整。如果需要查詢101學年度以前的紙本論文公開資訊,請聯繫圖資處紙本論文服務櫃台。如有不便之處敬請見諒。
開放時間 available 已公開 available

QR Code