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研究生: 陳正龍
Chan-Long Chen
論文名稱: 半導體量子點於癌細胞螢光標定之研究
Fluorescent Labeling of Cancer Cells with Semiconductor Quantum Dots
指導教授: 陳家俊
Chen, Chia-Chun
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2003
畢業學年度: 91
語文別: 中文
論文頁數: 69
中文關鍵詞: 量子點螢光標定奈米晶體半導體
英文關鍵詞: Quantum dot, fluorescent label, nanocrystal, semiconductor
論文種類: 學術論文
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  • 近年來,許多將無機奈米結構界面與生物系統相連的材料已引起細胞學及生物醫學上廣泛的應用。半導體量子點是目前研究細胞影像中最有希望的新興螢光標記物。在本篇論文中,我們利用水溶性硒化鎘/硫化鋅奈米晶體的螢光,標定人類乳癌細胞 ( MCF 7 ),以研究量子點進入細胞的方式、生物相容性、及在細胞內的光學穩定性。由共軛焦螢光顯微鏡的影像發現,這些細胞內的螢光訊號數天後依然穩定存在,而細胞亦不斷地在進行分裂。為了更進一步確認量子點是否能專一性地標定細胞內的特定胞器,我們將量子點的表面修飾上能辨識細胞核的胜肽以標定細胞核。實驗結果,大多數的螢光訊號對細胞核具有專一性而且比有機染料的螢光更強又穩定。這些實驗結果更促使未來能設計更具功能化的量子點以利活細胞內的蛋白質動態追蹤。

    Abstract
    Inorganic nanomaterials that interface with biological systems have
    recently attracted widespread interests in cellular and biomedical applications.
    Semiconductor quantum dots (QDs) are among the most promising emerging
    fluorescent labels for cellular imaging. In this thesis, to study the way into
    cell、biocompatibility and photostability of QDs, we fluorescently labeled
    breast cancer cells ( MCF 7 ) using water-soluble CdSe/ZnS core-shell
    nanocrystals. Confocal microscopy images indicated that the cells remained
    stably labeled for several days and continued cell division. However, to establish
    whether QDs can specifically label organelles at a subcellular level, we used
    QDs modified with the nucleus localization signal ( NLS ) peptides to label the
    nuclei of the cells. The results showed that much labeling signals are specific for
    the nuclei and are brighter and considerably more photostable than comparable
    organic dyes. These results encourage the construction of more complex
    functional QDs for protein tracking in cell.

    第一章 緒論.......................................1 1-1序言...........................................1 1-2半導體奈米材料.................................2 1-2-1 量子限量化效應...........................3 1-2-2 直遷能隙與非直遷能隙.....................5 1-2-3 量子點之基本性質及應用...................7 1-2-4 奈米晶體表面鈍化.........................10 1-3 細胞螢光技術之簡介............................11 1-4 細胞核及NLS peptide之功能.....................14 第二章 實驗.......................................17 2-1 研究動機與目的................................17 2-2 奈米晶體之合成................................18 2-2-1 合成CdSe、CdSe/ZnS奈米晶體所需藥品.......18 2-2-2 測量儀器.................................18 2-2-3 合成疏水性CdSe奈米晶體...................18 2-2-4 合成疏水性CdSe/ZnS奈米晶體...............20 2-2-5 合成親水性CdSe/ZnS奈米晶體...............20 2-3 固相peptide之合成.............................21 2-3-1合成NLS peptide序列所需之藥品.............21 2-3-2 儀器.....................................22 2-3-3 NLS peptide序列之合成步驟................23 2-4 合成生物官能化奈米晶體........................27 2-5 細胞培養......................................28 第三章 結果與討論.................................31 3-1 材料特性之分析................................31 3-1-1 疏水性硒化鎘/硫化鋅奈米晶體的性質探討....31 3-1-2 親水性硒化鎘/硫化鋅奈米晶體的性質探討....32 3-1-3 NLS 與 mNLS 之探討.......................33 3-2 硒化鎘/硫化鋅奈米晶體標定活細胞...............34 3-2-1 硒化鎘/硫化鋅奈米晶體進入細胞的過程探討..35 3-2-2 硒化鎘/硫化鋅奈米晶體的生物相容性........37 3-2-3 硒化鎘/硫化鋅奈米晶體在細胞內之光學性質..38 3-3 半導體奈米晶體辨識細胞核之探討................39 3-3-1 NLS 對奈米晶體辨識細胞核的影響...........39 3-3-2 m NLS 對奈米晶體辨識細胞核的影響.........41 3-3-3 不同大小的奈米晶體於螢光標定之探討.......42 3-3-4 螢光染劑的選擇...........................43 3-3-5 雷射掃描共軛焦影像切片分析...............44 第四章 結論與未來發展.............................64 參考文獻..........................................66

    1. Brus, L. E.; Efros, A. L.; Itoh, T., J. Lumin., 1996, 70, R7-R8.
    2. Weller, H., Philo. Trans. R. Soc. London, Ser. A, 1996, 354, 757.
    3. Ball, P. Nature, 2001, 414, 142.
    4. Kalyanasundaram, K.; Borgarello, E.; Duonghong, D.; Gratzel, M.,
    Angew. Chem. Int. Ed. Engl., 1981, 20, 987.
    5. Majetich, S. A.; Jin, Y., Science, 1999, 284, 470.
    6. Back, C. H.; Allenspach, R.; Weber, W.; Parkin, S. S. P.; Weller, D.;
    Garwin, E. L.; Siegmann, H. C., Science, 1999, 285, 864.
    7. Golden, A. N.; Echer, C. M.; Alivisatos, A. P., Science, 1992, 256, 1425.
    8. Wei, Q. H.; Bechinger, C.; Leiderer, P., Science, 2000, 287, 625.
    9. Wong, E. W.; Sheehan, P. E.; Lieber, C. M., Science, 1997, 277, 1971.
    10. Mitchell, P., Nature, 2001, 1013.
    11. Bruchez, M. Jr. Moronne, M., Gln. P., Weless, S. & Alivisatos, A. P.,
    Science, 1998, 281, 2013.
    12. Maria, E. A.; Warren, C. W. Chan.; Pirjo Laakkonen; Sangeeta, N.
    Bhatia.; Erkki Ruoslahti., Proc. Natl. Acad. Sci. USA, 2002, 99,
    12617.
    13. Jaiswal, J. K.; Mattoussi, H.; Mauro, J. M.; Simon, S. M.,
    Nat. Biotechnol, 2003, 21, 47.
    14.「電子材料」謝煜弘 編著,新文京開發出版有限公司.
    15. Alivisatos, A. P., Science, 1996, 271, 933.
    16. Wang, Y.; Herron, N., J. Phys. Chem. 1991, 95, 525.
    17.「固態電子學」呂助增 著,國立編譯館主編.
    18. Woggon, U., Springer, 1996, Berlin.
    19. Okada, P.; Lijima, S., Appl. Phys. Lett, 1991, 58, 1662.
    20. Danek, M.; Jensen, K. F.; Murray, C. B.; Bawendi, M. G., Appl. Phys.
    Lett., 1994, 65, 2795.
    21. Fujii, T.; Hisakawa, Y.; Winder, E. J.; Ellis, A. B., Bull. Chem. Soc.
    Jpn., 1995, 68, 1559.
    22. Gorer, S.; Hodes, G.; Sorek, Y.; Reisfeld, R., Materials Letters, 1997,
    31, 209.
    23. Grundemann, M. et al, Phys. Rev. Lett., 1995, 74, 4043.
    24. Petit, C.; Pileni, M. P., J. Phys. Chem., 1988, 92, 2282.
    25. Kortan, A. R.; Hull, R.; Opila, R. L.; Bawendi, M. G.; Steigerwald, M.
    L.; Carroll, P. J.; Brus, L. E., J. Am. Chem. Soc., 1990, 112, 1327.
    26. Steigerwald, M. L.; Alivisatos, A. P., Gibson, J. M.; Harris, T. D.;
    Kortan, R.; Muller, A. J.; Thayer, A. M.; Duncan, T. M.; Douglass,
    D. C.; Brus, L. E., J. Phys. Chem., 1988, 110, 3046.
    27.「光電工程-元件與原理」沈季燕 編著,全欣科技圖書.
    28. Huffaker, D. L.; Park, G.; Zou, Z.; Shchekin, O. B.; Deppe, D. G.,
    IEEE J. Selected Topics in Quantum Electron., 2000, 6, 452
    29. Peng, X., Schlamp, M. C., Kadavanich, A. V. & Alivisatos, A. P.,
    J. Am. Chem. Soc, 1997, 119, 7019.
    30. Murray, C. B., Norris, D. J. & Bawendi, M. G., J. Am. Chem. Soc,
    1993, 115, 8706.
    31. Dabbousi, B. O., Rodriguez-Viejo, J., Mikulec, F. V., Heine, J. R.,
    Mattoussi, H., Ober, R., Jensen, K. F. & Bawendi, M. G., J. Phys.
    Chem. B, 1997, 101, 9463.
    32. Hines, M. A.; Guyot-Sionnest, P., J. Phys. Chem. B, 1996, 100, 468.
    33. Manoj Nirmal and Louis Brus., Acc. Chem. Res. 1999, 32, 407.
    34.「生物電子顯微鏡學」陳家全、李家維、楊瑞森 著,行政院國科會精密儀器發展中心編印
    35. Peter, T. C.; Dong, C. Y.; Masters, B. R.; Berland, K. M.,
    Annu. Rev. Biomed. Eng., 2000, 2, 399.
    36. Cutler, S. R.; Ehrhardt, D. W.; Griffitts, J. S.; Somerville, C. R.,
    Proc. Natl. Acad. Sci. USA, 2000, 97, 3718.
    37. Conly, L. R.; Khodjakov, A., Science, 2003, 300, 91.
    38. Robert F. Service., Science, 1996, 272, 1423.
    39.「MOLECULAR CELL BIOLOGY」, Lodish、Berk、Zipursky、
    Matsudaira、Baltimore、Darnell, IV, 2000.
    40.「MOLECULAR BIOLOGY OF THE CELL」Alberts, B.; Bray, D.;
    Lewis, J.; Raff, M.; Roberts, K.; Watson J. D., III, 1994.
    41. Peng, X.; Schlamp, M. C.; Kadavanich, A. V.; Alivisatos, A. P.
    J. Am. Chem. Soc, 1997, 119, 7019.
    42. Mitchell, G. P.; Mirkin, C. A.; Letsinger, R. L., J. Am. Chem. Soc,
    1999, 121, 8122.
    43. Gerion, D.; Pinaud, F.; Williams, S. C.; Parak, W. J.; Zanchet, D.;
    Weiss, D. S.; Alivisatos, A. P., J. Phys. Chem. B, 2001, 105, 8861.
    44. Lanford, R. E.; Kanda, P.; Kennedy, R. C., Cell 1986, 46, 575.
    45. Merrifield, R. B., Science, 1986, 232, 341.
    46. Fischer-Fantuzzi, L. & Vesco, C., Mol. Cell. Biol, 1988, 8, 5495.
    47. Mattoussi, H.; Mauro, J. M.; Goldman, E. R.; Anderson, G. P.;
    Sundar, V. C.; Mikulec, F. V.; Bawendi, M. G., J. Am. Chem. Soc,
    2000, 122, 12142.
    48. Guo, w.; Li, J. J.; Wang, Y. A.; Peng, X., J. Am. Chem. Soc, 2003,
    125, 3901.
    49. Gaietta, G.; Deerinck, T. J.; Adams, S. R.; Bouwer, J.; Tour, O.;
    Laird, D. W.; Sosinsky, G. E.; Tsien, R. Y.; Ellisman., M. H.,
    Science, 2002, 296, 503.
    50. Miller, M. J.; Wei, S. H.; Parker, I.; Cahalan, M. D., Science, 2002,
    296, 1869.
    51. Wu, X.; Liu, H.; Liu, J.; Haley, K. N.; Treadway, J. A.; Larson, J. P.;
    Ge, N.; Peale, F.; Bruchez, M. P., Nat. Biotechnol, 2003, 21, 41.
    52. Parak, W. J. ; Boudreau, R.; Gros, M. L.; Gerion, D.; Zanchet, D.;
    Micheel, C. M.; Williams, S. C.; Alivisatos, A. P.; Larabell, C.,
    Adv. Mater. 2002, 14, 882.
    53. Winter, J. O.; Liu, T. Y.; Korgel, B. A.; Schmidt, C. E.,
    Adv. Mater. 2001, 13, 1673.
    54. Goldman, E. R.; Anderson, G. R.; Tran, P. T.; Mattoussi, H.
    Charles, P. T.; Mauro, J. M., Anal. Chem, 2002, 74, 841.
    55. Suh, j.; Writz, D.; Hanes, J., Proc. Natl. Acad. Sci. USA, 2003, 100,
    3879.
    56. Cartier, R.; Reszka, R., Gene Therapy, 2002, 9, 157.
    57. Duberter, B.; Skourides, P.; Norris, D. J.; Noireaux, V.; Brivanlou,
    A. H.; Libchaber, A., Science, 2002, 298, 1759.
    58. Eppstein, M. J.; Hawrysz, D. J.; Godavarty, A.; Sevick-Muraca, E. M.
    Proc. Natl. Acad. Sci. USA, 2002, 99, 9619.

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