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研究生： 賴昱宏
Lai, Yu-Hung
論文名稱： 雙邊不對稱球體自組裝行為之生物檢測平台
A Novel Biosensing Platform Based on the Self-assembly of Submicron Janus Particles
指導教授： 郭昌恕
Kuo, Chang-Shu
學位類別： 碩士
Master
系所名稱： 工學院 - 材料科學及工程學系
Department of Materials Science and Engineering
論文出版年： 2016
畢業學年度： 104
語文別： 英文
論文頁數： 91
中文關鍵詞： 雙邊不對稱球體 、專一性共同自組裝 、螢光共振能量轉移
外文關鍵詞： Janus particle, specific co-assembly, FRET
相關次數： 點閱：137  下載：0 
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    • 　　在本研究中，合成次微米雙邊不對稱球體進行具有專一性球體的共同自組裝，並進一步結合螢光共振能量轉移做為一生物傳感器機制。共同自組裝行為的主要材料為雙邊不對稱球體，是以五百奈米的二氧化矽球體為主體，對其中一個半球表面進行氨基的改質，並依序接上Marina Blue 螢光染料及抗生物素抗體。共同自組裝行為中，帶有負電荷球體為市售之聚丙烯腈奈米微珠，其粒徑為八十奈米且帶有羧酸化的表面，並已經預先帶有螢光染料Chromeon 470在其中。在聚丙烯腈奈米微珠表面接上生物素為抗原做後續研究使用。Marina Blue螢光染料扮演的是螢光共振能量轉移的供體物質，而Chromeon 470則為螢光共振能量轉移的受體物質。當供體物質與受體物質距離很近的時候會有螢光共振能量轉移的發生，藉由偵測到螢光共振能量轉移現象，可以證實球體共同自組裝聚合體的生成。
    　　表面帶有胺基之雙邊不對稱球體依循過去研究發展所製備。在球體共同自組裝後，利用動態光散射粒徑分析儀進行研究並監測。理想之聚合體是由四顆五百奈米雙邊不對稱球體與一顆八十奈米聚丙烯腈奈米微珠共同自組裝組成的四面體。當球體共同自組裝成聚合體，可即時以動態光散射粒徑分析儀觀測其粒徑大小的增加，及偵測螢光共振能量轉移放射光之光致發光訊號。藉由改變兩種球體的數量比例，共同自組裝聚合體可形成不同的形態，如：釋迦頭狀結構、二聚體、三聚體、由四顆球組成的四面體、由五顆球組成的六面體等。結合穩定的球體共同自組裝及螢光共振能量轉移效應，可實現其作為一新穎的生物傳感器機制和平台。

    In this research work, submicron Janus particles were synthesized for specific particle co-assembly, which further triggered the Förster resonance energy transfer (FRET) as the biosensing mechanism. The major material of particle co-assembly included Janus particles cored with 500 nm silica particles and functionalized with amino-silane in one of their hemispheric surfaces, followed by their conjugation with Marina Blue dye and anti-biotin antibody. The anionic particles for the co-assembly were commercially-available Chromeon 470 particles (Ch470), which were 80 nm polyacrylonitrile (PAN) particles with carboxylated surface and contained Chromeon 470 dye. Ch470 particle surfaces were also conjugated with biotin as antigen before the co-assembly studies. Marina Blue and Chromeon 470 were utilized as the FRET donor and acceptor, respectively. Since the FRET were triggered only when the distance between the donor and acceptor was very close, the successful FRET signals also revealed the cluster formation from the particle co-assembly.
    The preparation of amino Janus particles followed the prior synthetic approach developed in the prior work. The particle co-assembly was investigated and monitored using the dynamic light scattering (DLS). The ideal assembled clusters were the so-called tetramers that four 500 nm Janus particles assembled one 80 nm Ch470 particles. Well-defined particle co-assembly simultaneously initiated the cluster diameter increase in the DLS measurement, and the FRET emission from their photoluminescence outputs. Depended on the particle ratios, the assembled particle clusters also included the morphologies of “Buddha-head”, Dimer, trimer, tetramer (tetrahedron) and pentamer (hexahedron). Steady particle co-assembly and the related FRET responses realized the novel biosensing mechanism and platform.

    中文摘要 I Abstract II 致謝 III Table of contents IV List of Illustrations VI List of Table IX Chapter 1 Introduction 1 1.1. Janus particles and asymmetric micro- and nano- materials 1 1.1.1. Janus-like Materials 2 1.1.2. Fabrications of Janus particles 3 1.1.3. One-dimensional electrospun polymer fibers as particle embedding substrates 12 1.1.4. Applications 14 1.2. Particle Self-assembly 18 1.2.1. Introduction to self-assembly 18 1.2.2. Field-induced Self-assembly 24 1.2.3. Applications of self-assembly 25 1.3. Förster resonance energy transfer 27 1.3.1. Mechanism 28 1.3.2. Applications 31 1.4. Biosensor 33 Chapter 2 Research Motivations 34 Chapter 3 Experimental 35 3.1. Chemical 35 3.2. Instruments Used in the Material Fabrication 38 3.3. Experiment process－Amine-modified Janus particles 39 3.3.1. PMMA/P4VP solution preparation 39 3.3.2. Electrospinning process 39 3.3.3. 500 nm silica colloidal solution preparation 40 3.3.4. Dipping process- colloid adsorption 40 3.3.5. Heat treatment process- embedding 41 3.3.6. APS modification process- CVD 41 3.3.7. Polymer fibers dissolving process 42 3.3.8. Dye graft process 42 3.3.9. Antibody modify 43 3.3.10. Functionalization of Chromeon 470 particles 43 3.4. Analytical Instruments 44 3.4.1. Particle size distribution (dynamic light scattering) 44 3.4.2. Zeta potential 44 3.4.3. Dark field optical microscopy 45 3.4.4. UV-vis spectrometer (UV-vis) 45 3.4.5. Photoluminescence spectroscopy (PL) 46 3.4.6. Scanning electron microscopy (SEM) 46 3.4.7. Photoluminescence optical system 47 Chapter 4 Results and Discussion 48 4.1. Characterizations of Janus Particles and FRET dyes 48 4.1.1. CVD APS amino group quantification 48 4.1.2. Marina Blue, Chromeon 470 and their FRET 50 4.1.3. Antibody-Janus particle conjugation 52 4.1.4. Biotin-Ch470 conjugation 54 4.2. The co-assembly behavior of Janus particles and 100 nm particles 56 4.2.1. Electrostatic interaction and reversible dynamic co-assembly 56 4.2.2. Specific interaction 67 4.3. FRET effect of specific co-assembly of Janus particle 73 4.3.1. Specific interaction co-assembly and FRET effect 73 4.3.2. Irreversible specific assembly 79 Chapter 5 Conclusion 82 Chapter 6 Reference 83

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