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論文名稱 Title |
以能量轉移機制來發展半導體高分子奈米顆粒,並應用於分析感測器上 Applications of Semiconducting Polymer Dots in Sensing via FRET Strategy |
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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 |
110 |
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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 |
2014-06-23 |
繳交日期 Date of Submission |
2014-07-28 |
關鍵字 Keywords |
螢光能量共振轉移、光控分子、螢光訊號比例、半導體高分子奈米顆粒、比色法、聚二乙炔、近紅外光、銅離子 Förster resonance energy transfer, Bioimaging, Polydiacetylene, Near-infrared, Fluorescent probe, Spiropyran, merocyanine, ratiometric, Semiconducting Polymer Dots |
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統計 Statistics |
本論文已被瀏覽 5650 次,被下載 1161 次 The thesis/dissertation has been browsed 5650 times, has been downloaded 1161 times. |
中文摘要 |
本篇研究利用半導體高分子奈米顆粒(Semiconducting Polymer Dots,Pdots)具良好的光學特性,並且含高能量傳遞的優點來設計分析感測器。除此之外,結合FRET (Förster resonance energy transfer)能量傳遞發展以半導體高分子奈米顆粒為主體做不同的探討與應用,在此作的應用分別為金屬離子的偵測,以及製備具近紅外光特性的螢光探針。 (一)修飾上光控分子的半導體高分子奈米顆粒以螢光訊號比例的方式,對銅離子做偵測: 銅離子是人體含量前三多的金屬離子之一,含量過多會導致肝硬化,嚴重甚至是死亡,但含量過少則會引發貧血、骨質酥鬆等症狀,所以銅離子的偵測非常重要。在此研究中,我們分別選擇PPE和PFBT這兩種Pdots,利用兩者之間能產生有效的FRET能量傳遞,並在Pdots上修飾可以抓取銅離子的光控分子(spiropyran,SP),SP為一種光控分子,照射UV光源時會形成merocyanine (MC)結構,而MC照射白光則會變回SP。另外,以SP (spiropyran)上的COOH和CO520鍵結合成SP-CO520,然後以再沉澱法和Pdots結合,由於CO520的結構為一端疏水性一端親水性,所以合成的SP-CO520-PPE和SP-CO520-PFBT,就會以CO520的疏水端部分往Pdots內部,而CO520上的PEG為親水端,則迫使SP在Pdots外部。由於SP結構上的碳鍊長度所造成的空間大小最吻合銅離子,所以只需要簡單改變光源即可對銅離子具選擇性偵測,除此之外,以螢光訊號比例的方式作探討,設計出簡單又方便的偵測系統。 (二)發展polydiacetylene包覆近紅外光半導體高分子奈米顆粒,並應用在生物影像以及感測器: 由於Near-infrared (NIR,650 nm-900 nm) 放光具有高穿透性、對生物體傷害小、低背景干擾等優點,所以為科學家目前致力發展的目標。本研究利用PFBT-DBT Pdots包覆NIR染劑,使PFBT-DBT透過FRET傳能量給NIR染劑,而讓Pdots放光由650 nm改變到NIR範圍。首先,測試九種不同NIR 染劑,得到FRET效果較好的三個染劑 (NIR653、NIR695、NIR775),再由螢光光譜結果,選擇NIR695及NIR775去做進一步探討,分別藉由有無包覆polydiacetylene (聚二乙炔,PDA) 以及浸泡至37℃來測試穩定性。結果顯示,有PDA包覆,使NIR-emitting Pdots在37℃下浸泡24小時,NIR染劑還是良好存在Pdots中。後續,使用HeLa cell (子宮頸癌細胞) 進行內吞作用,可得知在酸性狀態下不會破壞螢光訊號,此外,利用PDA上的COOH官能基來進行bioconjugation,成功標記在MCF-7 cell (乳癌細胞) 表面。最後,利用PDA具有比色法 (colorimetry) 的特性來製備試紙,原本呈現藍色的試紙在90oC則變成紅色。綜合以上,本研究具備了螢光以及比色法 (colorimetry) 的雙偵測優勢。 |
Abstract |
1.Photoactivated ratiometric copper(II) ion sensing with semiconducting polymer dots This paper describes a simple platform that employs spiropyran-functionalized semiconducting polymer dots as a fluorescent probe for photoactivated ratiometric and sensitive Cu2+ detection, in which the sensing mechanism is based on photogenerated merocyanine that can selectively bind Cu2+ to induce Förster resonance energy transfer. 2. Polydiacetylene-Enclosed Near-Infrared Fluorescent Semiconducting Polymer Dots for Bioimaging and Sensing Semiconducting polymer dots (Pdots) recently have emerged as a new type of ultrabright fluorescent probe with promising applications in biological imaging and detection. With the increasing desire for near-infrared (NIR) fluorescing probes for in vivo biological measurements, the currently available NIR-emitting Pdots are very limited and the leaching of the encapsulated dyes/polymers has usually been a concern. To address this challenge, we first embedded the NIR dyes into the matrix of poly [(9, 9 -dioctylfluorene)-co-2,1,3-benzothiadiazole-co-4,7-di(thiophen-2-yl)-2,1,3-benzothiadiazole] (PFBT-DBT) polymer and then enclosed the doped Pdots with polydiacetylenes (PDAs) to avoid potential leakage of the entrapped NIR dyes from the Pdot matrix. These PDA-enclosed NIR-emitting Pdots not only emitted much stronger NIR fluorescence than conventional organic molecules but also exhibited enhanced photostability over CdTe quantum dots, free NIR dyes, and gold nanoclusters. We next conjugated biomolecules onto the surface of the resulting Pdots and demonstrated their capability for specific cellular labeling without any noticeable nonspecific binding. To employ this new class of material as a facile sensing platform, an easy-to-prepare test paper, obtained by soaking the paper into the PDA-enclosed NIR emitting Pdot solution, was used to sense external stimuli such as ions, temperature, or pH, depending on the surface functionalization of PDAs. We believe these PDA-coated NIR-fluorescing Pdots will be very useful in a variety of bioimaging and analytical applications. |
目次 Table of Contents |
誌謝…………………………………………………………………...…………..……..i 中文摘要………………………………………….…………………..……………..….ii 英文摘要……………………………………………………………..…………..…….iv 目錄………………………………………………………………….…….…...………vi 圖目錄……………………………………………………………….…….…...………ix 表目錄……………………………………………………………….………...…..…...xv 縮寫表……………………………………………………………….………...…..…...xv 第一章、 緒論………………………………………………………………………….1 一、前言……………………………………………………………………….………...1 1-1.歷史發展…………………………………………………………………….………1 1-2.製備方式………………………………………………………………….………..11 1-3.應用………………………………………………………………………….……..12 1-3-1.離子偵測……………………………………………………….….......………..13 1-3-2.生物偵測……………………………………………………….….………........14 1-3-3.生物影像………………………………………………………………….......…14 1-4.螢光共振能量轉移 (Förster resonance energy transfer,FRET)…….......... 15 二、研究動機…………………………………………………………………………..20 第二章、半導體高分子奈米顆粒以光活化且螢光訊號比例的方式,對銅離子做偵測 一、前言………………………………….………………………………...…..……….21 二、實驗部分 2-1.實驗藥品……………………………………….……………………………...…...22 2-2.儀器裝置…………………………………………………………………………...24 2-3.樣品配製方法……………………………………………………………………...25 2-4.實驗步驟…………………………………………………………………………...26 三、實驗設計及原理…………………………………………………………………..29 四、實驗結果與討論 4-1.偵測系統的檢測…………………………………………………………………...32 4-2.半導體高分子奈米顆粒的尺寸…………………………………………………...34 4-3.以螢光檢測不同銅離子濃度……………………………………………………...36 4-4.選擇性……………………………………………………………………………...38 4-5.偵測系統的可逆性研究…………………………………………………………...39 4-6. Förster radius及FRET效率的計算………………………………………………40 4-7.真實樣品…………………………………………………………………………...42 五、結論………………………………………………………………………………..42 第三章、發展polydiacetylene包覆近紅外光半導體高分子奈米顆粒,並應用在生物 影像以及感測器 一、 前言……………………………………………………………………….……....43 二、 實驗部分 2-1.實驗藥品………………………………………………………….……….……….47 2-2.儀器裝置…………………………………………………………………….……..50 2-3.樣品配製方法……………………………………………………………….….….52 2-4.實驗步驟………………………….………………………………………….….…54 三、實驗設計及原理………………………………………………………….….……60 四、實驗結果與討論 4-1.選擇半導體高分子奈米顆粒以及NIR染劑……………………………………....62 4-2. PFBT-DBT與NIR染劑的光學性質及奈米顆粒尺寸 ...................................64 4-3.探討NIR染劑的含量................................................................................66 4-4. Polydiacetylene的包覆效率....................................................................68 4-5.修飾上diacetylene的NIR半導體高分子奈米顆粒之穩定性測試....................69 4-6.螢光量子產率的探討...............................................................................73 4-7. Förster radius及FRET效率的計算 ...........................................................76 4-8. NIR695-PFBT-DBT半導體高分子奈米顆粒的螢光生命期...........................78 4-9. NIR695-PFBT-DBT半導體高分子奈米顆粒的光穩定性...............................79 4-10.PDA包覆0.75% NIR695-PFBT-DBT半導體高分子奈米顆粒在生物體系的應用 ....................................................................................................................80 4-10-1. 細胞內吞作用 (Endocytosis)...........................................................80 4-10-2. 細胞特異性標記 (Specific cellular targeting).....................................81 4-10-3. 製備偵測型試紙 83 五、結論……………………………………….……………. ..….……………...……84 第四章、參考文獻……………………………………….…….….……………...…...85 |
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