雷射環狀模態及螢光生命週期於雙光子激發受激輻射耗損顯微術解析度提升之研究

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姓名

李書恩(Shu-En Li) 查詢紙本館藏

畢業系所

照明與顯示科技研究所

論文名稱

雷射環狀模態及螢光生命週期於雙光子激發受激輻射耗損顯微術解析度提升之研究
(The effects of donut mode and fluorescence lifetime on the resolution improvement of two-photon excitation STED microscopy)

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摘要(中)

本論文是以雙光子激發受激輻射耗損顯微鏡為目標，討論環形光場分布以及樣品螢光生命週期對其解析度之影響，提出增加解析度的方法。在環形光場的討論中，本研究採用模擬及光場量測分析以不同方式所產生的環形光場及其在強聚焦下的光場變化，而螢光生命週期的研究則是模擬的方式來了解螢光樣品的選擇可以如何幫助解析度的提昇。此外，本研究已著手進行系統的建構，期望能透過實際系統的建構來證實上述論點之可行性。

摘要(英)

This thesis is based on the two-photon excitation stimulated-emission depletion microscopy to discuss the effects of donut beam and fluorescence lifetime on the spatial resolution of this system and propose the methods to improve the resolution. In this research, both simulation and experimental measurements were applied to analyze the intensity distributions of donut beams generated in different ways and their distributions under tightly-focusing condition. On the other hand, only simulation was used to discuss the impact of fluorescence lifetime on the spatial resolution improvement. Finally, in order to obtain the experimental evidences of our results, the two-photon excitation stimulated-emission depletion microscopy was setup.

關鍵字(中)

★ 顯微術
★ 螢光
★ 受激輻射耗損顯微術
★ 環形光場

關鍵字(英)

★ microscopy
★ STED microscopy
★ fluorescence
★ donut beam

論文目次

目錄
頁次
摘要...................................i
Abstract...................................ii
致謝...................................iii
目錄...................................v
圖目錄...................................vii
第一章導論...................................1
1-1 本文緣起...................................1
1-2 論文大綱...................................2
第二章光學顯微術...................................4
2-1 光學顯微術簡述...................................4
2-2 光學顯微術成像原理...................................5
2-3 各種超解析顯微術簡介...................................7
第三章受激輻射耗損顯微術...................................10
3-1 螢光發光原理.......................................................................................10
3-2 螢光光學顯微術...................................................................................12
3-3 受激輻射耗損顯微術...........................................................................15
3-3-1 原理簡介.....................................................................................15
3-3-2 各種STED技術介紹.................................................................18
3-3-3 受激輻射耗損效率.....................................................................19
第四章 STED解析度之討論……....................................................................22
4-1 前言.......................................................................................................22
4-2 環形光場...............................................................................................22
4-2-1 高數值孔徑聚焦下光場分布之討論.........................................23
4-2-2 環形光場產生方法及測量.........................................................26
4-2-3 環形光場之比較.........................................................................34
4-3 螢光生命週期v.s.時間閘受激輻射耗損顯微術.................................37
4-3-1 時間閘受激輻射耗損顯微術.....................................................37
4-3-2 樣品生命週期之選擇.................................................................40
4-4 雙光子受激輻射耗損顯微鏡之建構...................................................42
4-4-1 雙光子螢光顯微鏡.....................................................................42
4-4-2 受激輻射耗損效率之量測….....................................................44
4-4-3 反掃描系統及 STED 影像測試..............................................50
第五章結論與未來展望...................................................................................52
參考文獻.............................................................................................................53

參考文獻

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指導教授

陳思妤(Szu-Yu Chen)

審核日期

2013-7-11

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