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論文名稱 Title |
利用HBT 光子相干技術及單分子螢光量測系統來測量 DiI 分子的光物理特性 Single molecule fluorescence and Hanbury Brown-Twiss photon-correlation technologies study DiI molecule |
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系所名稱 Department |
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畢業學年期 Year, semester |
語文別 Language |
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學位類別 Degree |
頁數 Number of pages |
70 |
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研究生 Author |
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指導教授 Advisor |
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召集委員 Convenor |
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口試委員 Advisory Committee |
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口試日期 Date of Exam |
2006-06-30 |
繳交日期 Date of Submission |
2006-07-16 |
關鍵字 Keywords |
螢光生命期、單分子、螢光強度、光子反成串 photon anti-bunching, fluorescence lifetime, single molecule, fluorescence intensity |
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統計 Statistics |
本論文已被瀏覽 5658 次,被下載 0 次 The thesis/dissertation has been browsed 5658 times, has been downloaded 0 times. |
中文摘要 |
我們建構一套單分子螢光量測系統,具有同時測量分子的瞬時螢光強度(transient fluorescence intensity)、螢光生命週期( fluorescence lifetime )、螢光偏振方向(fluorescence anisotropy)、以及使用Hanbury Brown-Twiss 光子相干技術來觀察單一DiI (1,1'-dioctadecyl- 3,3,3',3'- tetramethylindocarbocyanine perchlorate )分子的反成串(anti-bunching)行為等光物理參數,並應用於測量DiI染料分子的光物理特性。 Cyanine染料是指包含兩個氮原子為主之芳香族集團,而中間以共軛鍵結連接而成之有機分子。這類分子在光致敏(photosensitizer)、光儲存媒介、非線性光學材料、雷射染料、以及生物分子之螢光標定等許多方面具有優良性質而廣受注目。其中,由於DiI 分子(cyanine dyes 中的一員)在固態媒介環境下具有良好的光穩定(photo-stability)、高量子效率(quantum yield)以及較低的躍遷至三重態機率(intersystem crossing rate)等特性,使得DiI在單分子螢光量測中一直都扮演著重要的角色。 我們的實驗目標是藉著比較不同光物理參數間之關聯來瞭解所觀測的分子特性。我們有得到典型的DiI分子結果,類似一般所知的研究結果,具有清楚的on-off blinking行為,Fluorescence anti-bunching表現,one-step photo-bleaching行為,以及一致的polarization方向。除此之外,某些分子在觀測時具有部分參數之變化,顯示觀測過程中分子結構的變化。而且有些觀測點則表現螢光光子間距時間趨近於零時其出現機率不為零,這顯示觀測點上存在著超過一個分子的結構。實驗結果顯示,藉著這些參數的分析,我們可以有效的分辨不同分子的特性。 |
Abstract |
We have constructed a single molecule detection system with the capability to simultaneously measure many parameters, including transient fluorescence intensity, fluorescence lifetime, and photon anti-bunching behavior via the Hanbury Brown-Twiss photon-correlation technique. In addition, we apply the system to study the single DiI (1, 1 '- dioctadecyl- 3, 3 , 3 ', 3 ' - tetramethylindocarbocyanine perchlorate) molecule, to characterize the photo-physical behaviors. Cyanine dyes are the molecules that constitute of two nitrogen centers, one of which is positive charged, and is linked by a conjugated chain with odd number of carbon atoms to the other nitrogen center. Cyanine dyes are interested in the photo sensitization, optical recording media, nonlinear optics, laser dyes, and many interesting photophysical and photochemical behaviors. Among them, DiI plays an important role in single molecule fluorescence investigations. The high photo-stability, good QE, and low inter-system crossing rates, make it a pioneer for the widely investigations in single molecule studies. Our experimental goal is to understand the characteristic of the monitored single molecule by the measuring photo-physical parameters. Our results include the typical behaviors in DiI molecules: clear on-off blinking, fluorescence anti-bunching, one-step photo-bleaching, and consistent fluorescence polarization orientation. In addition, we also observed some change during measurement, which indicates the corresponding change of structure. Few molecules also exhibit non-zero probability around the zero delay time, which indicates the simultaneous existence of more than one quantum emitters in the detected region. These results demonstrate that the parameters are essential for understanding and characterizing the observed molecules in single molecule level. |
目次 Table of Contents |
目錄 頁碼 中文摘要...................................................................................................ii 英文摘要..................................................................................................iii 誌謝...........................................................................................................v 目錄..........................................................................................................vi 圖目錄....................................................................................................viii 第一章 緒論.......................................................................................1 第二章 原理.......................................................................................5 2.1 節 : 螢光及磷光簡介 .................................................5 2.2 節 : 樣品簡介.............................................................10 2.3 節 : 螢光量測架構.....................................................11 2.4 節: Time-Correlated Single Photon Counting (TCSPC)………………………………………...15 2.5 節 : Hanbury Brown-Twiss setup 簡介.......................18 2.6 節 : 螢光極化(polarization)方向的偵測....................19 第三章 實驗架構及步驟..................................................................21 3.1 節 : 儀器架設..............................................................21 3.2 節 : 樣品準備..............................................................26 3.3 節 : 實驗步驟..............................................................27 第四章 實驗結果與討論..................................................................35 4.1 節 : 眾多分子之 Anti-bunching 行為......................35 4.2 節 : 典型單一DiI分子的螢光強度與時間關係…..36 4.3 節 : 非典型單一DiI分子的螢光強度與 時間的關係..........................................................42 第五章 結論與未來展望.................................................................56 參考文獻.................................................................................................59 |
參考文獻 References |
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