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系統識別號 U0026-0812200915273737
論文名稱(中文) 無孔徑近場掃描式顯微術於螢光影像之研究
論文名稱(英文) Apertureless near-field scanning microscopy for fluorescence imaging
校院名稱 成功大學
系所名稱(中) 工程科學系碩博士班
系所名稱(英) Department of Engineering Science
學年度 97
學期 2
出版年 98
研究生(中文) 陳長營
研究生(英文) Chang-Ying Chen
電子信箱 n9696436@mail.ncku.edu.tw
學號 n9696436
學位類別 碩士
語文別 中文
論文頁數 85頁
口試委員 指導教授-陳顯禎
口試委員-張世慧
口試委員-羅裕龍
中文關鍵字 信號調變  奈米結構  螢光信號  無孔徑近場掃描式光學顯微術 
英文關鍵字 fluorescent signal  nanostructure  signal modulation.  apertureless near-field scanning optical microsc 
學科別分類
中文摘要 傳統的光纖式近場光學顯微鏡受到截止效應、熱傷害等問題影響,造成解析度有所限制,因此可開發無孔徑的近場光學掃描式顯微鏡(apertureless near-field scanning optical microscope,aNSOM)來突破。其原理主要藉著原子力顯微鏡(atomic force microscope,AFM)的探針與入射光場作用,產生區域電場強化的效應,與樣品作用後產生一有效散射場。論文主要的研究是以AFM為基礎,研製一套aNSOM,期望其光學空間解析度可達到10nm以下,並將其應用於螢光樣品的影像量測。
研發之aNSOM由於其收光的顯微物鏡位於遠場偵測干涉後的高頻干涉信號,其信號包含針尖與奈米結構作用後的近場光學信號及遠場的背景雜訊,為了增加影像的訊噪比(signal-to-noise ratio,SNR),利用距離調變技術且搭配自差式干涉(homodyne interferometry)與外差式干涉(heterodyne interferometry)系統擷取信號;其中外差式干涉技術多引入一道可控之參考信號,對於影像之SNR之提昇比自差式干涉更有效。另外,論文中也針對近場螢光影像作相關之討論與量測,並分析探針與螢光分子的電場交互作用及螢光信號的擷取,期望未來可將本系統應用於分子生物學,得到分子尺度的醫學螢光影像。
英文摘要 The optical spatial resolution of conventional aperture near-field scanning optical microscopy is limited by cut-off effect and thermal noise effects. An apertureless near-field scanning optical microscope (aNSOM) has been developed to break the limitation. The localized electromagnetic field near the region between the sample and the tapping tip of atomic force microscopy (AFM), and then the effective scattering field which including near-field optical signal will be detected by far-field microscopy. This thesis attempts to develop an aNSOM based on a commercial AFM to achieve an optical spatial resolution better than 10 nm. Furthermore, the aNSOM is applied to measure near-field fluorescence signal
In the system, the modulated optical signal including the near-field signal and background noise is collected by a far-field microscope. In order to improve the near-field image with a high signal-to-noise ratio (SNR), the signal from the tapping tip at high harmonic terms is demodulated for both homodyne and heterodyne interferometric detections. With the help of an addition reference signal, the heterodyne detection can improve the SNR by rejecting near-field background noise. To test the system performance, a near-field fluorescent image from fluorescence molecules is grabbed. The interaction between the fluorescence signal and the tip is studied to assist how to obtain the fluorescent signal more efficiency. Eventually, the system could play a key role in biomolecular field, and provide fluorescence image with a molecular -scale resolution.
論文目次 摘要......................................................I
Abstract.................................................II
誌謝.....................................................IV
目錄......................................................V
圖目錄.................................................VIII
表目錄...................................................XI
第一章 緒論...............................................1
1-1 前言..................................................1
1-2 研究動機及目的........................................2
1-3 文獻回顧..............................................3
1-4 論文架構..............................................6
第二章 無孔徑近場掃描式光學顯微術.........................7
2-1 光學空間解析度........................................7
2-2 原子力顯微鏡之原理與系統.............................11
2-3 無孔徑近場掃描式光學顯微術之原理.....................14
2-3-1 外加光場與表面分子之交互作用......................15
2-3-2 區域性表面電漿子共振..............................16
2-3-3 入射場與探針尖端粒子之電磁關係....................17
2-3-4 探針尖端粒子與樣品表面之電磁關係..................19
2-3-5 探針形態與強化因子之關係..........................22
2-4 調變技術.............................................24
2-4-1 距離調變..........................................24
2-4-2 自差式與外差式干涉原理............................25
第三章 近場螢光信號號....................................31
3-1 螢光之原理...........................................31
3-1-1 原子能階與螢光....................................32
3-1-2 吸收光譜與發射光譜................................33
3-1-3 螢光之生命周期、發光效率、淬滅....................34
3-2近場螢光顯微術........................................37
3-2-1 探針與螢光分子之交互作用..........................37
3-2-2 近場螢光影像......................................39
第四章 系統架構..........................................41
4-1 光學系統設計.........................................41
4-2 機構設計.............................................48
4-3 硬體設備與電路設計...................................51
4-3-1 硬體設備..........................................51
4-3-2 電路設計..........................................56
4-4 信號擷取及控制流程...................................58
第五章 實驗結果與討論....................................62
5-1 系統校正與樣品量測測試...............................62
5-1-1自差式干涉術之量測分析.............................64
5-1-2外差式干涉術之量測分析.............................68
5-2 螢光分子影像.........................................71
5-2-1 螢光樣品製備......................................71
5-2-2 螢光影像分析......................................72
第六章 結論與未來工作....................................78
參考文獻.................................................80
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