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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 朱士維(Shi-Wei Chu) | |
dc.contributor.author | Kai-Wen Lai | en |
dc.contributor.author | 賴楷文 | zh_TW |
dc.date.accessioned | 2021-06-15T01:13:52Z | - |
dc.date.available | 2014-08-19 | |
dc.date.copyright | 2011-08-19 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-16 | |
dc.identifier.citation | 1. S. W. Hell, 'Far-field Optical Nanoscopy', Science 316, 1153 (2007).
2. E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. L. Schwartz, and H. F. Hess, 'Imaging Intracellular Fluorescent Proteins at Nanometer Resolution', Science 313, 1642 (2006). 3. M. J. Rust, M. Bates, and X. Zhuang, 'Sub-diffraction-limit Imaging by Stochastic Optical Reconstruction Microscopy (STORM),' National Methods 3, 793 (2006) 4. S. W. Hell and J. Wichmann, 'Breaking the Diffraction Resolution Limit by Stimulated Emission: Stimulated-Emission-Depletion Fluorescence Microscopy', Optical Letters 19, 780 (1994). 5. E. Rittweger, K. Y. Han, S. E. Irvine, C. Eggeling, and S. W. Hell, 'STED Microscopy Reveals Crystal Colour Centres with Nanometric Resolution', Nature Photonics 3, 144 (2009). 6. J. K. Ranka, R. S. Windeler, and A. J. Stentz, 'Visible Continuum Generation in Air-silica Microstructure Optical Fibers with Anomalous Dispersion at 800 nm', Optical Letters 25, 25 (2000). 7. P. Russell, 'Photonic Crystal Fibers', Science 299, 358 (2003). 8. S. W. Hell, D. Wildanger, E. Rittweger, and L. Kastrup, 'STED Microscopy with a Supercontinuum Laser Source', Optics Express 16, 9614 (2008). 9. E. Hecht, 'Optics ', 4th Ed, (Addison Wesley, USA, 2002). 10. M. Gu, 'Principles of Three Dimensional Imaging in Confocal Microscopes', (World Scientific, Singapore, 1996). 11. N. S. Claxton, T. J. Fellers, and M. W. Davidson, 'Laser Scanning Confocal Microscopy', http://www.olympusmicro.com. 12. B. Valeur, 'Molecular Fluorescence: Principles and Applications', (Wiley-VCH, USA, 2002). 13. D. J. Griffiths, 'Introduction to Quantum Mechanics', (Pearson Prentice Hall, USA, 2005). 14. S. W. Hell, M. Dyba, and S. Jakobs, 'Concepts for Nanoscale Resolution in Fluorescence Microscopy', Current Opinion in Neurobiology 14, 599 (2004). 15. S. W. Hell, A. Schönle, and A. Bos, 'Nanoscale Resolution in Far-Field Fluorescence Microscopy', in Science of Microscopy, P. W. Hawkes, J. C. H. Spence, Editors (Springer, USA, 2007), 790-834. 16. V. Westphal and S. W. Hell, ' Nanoscale Resolution in the Focal Plane of an Optical Microscope', Physical Review Letter 94, 143903 (2005). 17. E. Rittweger, B. R. Rankin, V. Westphal, and S. W. Hell, 'Fluorescence Depletion Mechanisms in Super-resolving STED microscopy', Chemical Physics Letters 442, 483 (2007). 18. K. I. Willig, S. O. Rizzoli, V. Westphal, R. Jahn, and S. W. Hell, 'STED Microscopy Reveals that Synaptotagmin Remains Clustered after Synaptic Vesicle Exocytosis', Nature 440, 935 (2006). 19. J. Keller, A. Schonle, S. W. Hell, 'Efficient Fluorescence Inhibition Patterns for RESOLFT Microscopy', Optics Express 15, 3361 (2007). 20. B. R. Boruah and M. A. A. Neil, 'Susceptibility to and Correction of Azimuthal Aberrations in Singular Light Beams', Optics Express 14, 10377 (2006). 21. S. Schrof, T. Staudt, E. Rittweger, N. Wittenmayer, T. Dresbach, J. Engelhardt, and S. W. Hell, 'STED Nanoscopy with Mass-produced Laser Diodes', Optics Express 19, 8066 (2011). 22. G. Moneron, R. Medda, B. Hein, A. Giske, V. Westphal, and S. W. Hell, 'Fast STED Microscopy with Continuous Wave Fiber Lasers', Optics Express 18, 1302 (2010). 23. Kenneth R. Spring, Thomas J. Fellers, and Michael W. Davidson, “Confocal Microscope Scanning Systems”, http://www.olympusmicro.com. 24. V. Westphal, M. A. Lauterbach, A. Di Nicola, S. W. Hell, 'Dynamic Far-field Fluorescence Nanoscopy', New Journal of Physics 9, 435 (2007) 25. D. Wildanger, R. Medda, L. Kastrup, S. W. Hell, 'A Compact STED Microscope Providing 3D Nanoscale Resolution', Journal of Microscopy 236, 35(2009). | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42443 | - |
dc.description.abstract | 螢光顯微鏡結合共軛焦顯微鏡成為研究生物分子影像最普遍的技術,因為比起電子顯微鏡,樣本的備製較為容易,也提供非侵入式的觀測,並且利用共軛焦顯微鏡光學切片的能力來取得三維影像。然而,解析度受到光學的繞射極限限制,解析度大約為所使用的波長的一半。近十幾年來,許多超高解析度的光學顯微技術被研發出來,其中受激發射耗伐顯微術在使用一般螢光染劑可達到二十至八十奈米的解析度,因此材料科學與生醫影像得以突破繞射極限到達奈米尺度的研究。
本篇論文中主要的研究是以商用的雷射掃描共軛焦顯微鏡為基礎架設受激發射耗乏顯微鏡,並且以超連續雷射做為光源。解析度的提升已經由自製的受激發射耗乏顯微鏡達成,但要達到更好的解析度,需要解決許多系統內在的像差問題。超連續雷射光源寬闊的波長範圍可提供廣泛的螢光分子應用。 | zh_TW |
dc.description.abstract | Fluorescence microscopy combined with confocal laser scanning microscope is the most common way for imaging bio-molecules. Since the easier sample preparation compared with electron microscopy, non-invasive investigation and 3D image acquisition are expected. However, the resolution of optical microscopy is limited by diffraction barrier, which is about half of incident wavelength. In the past ten years, many super-resolution techniques are successfully developed. Stimulated emission depletion (STED) microscopy is a kind of super-resolution microscopy, the resolution can be enhanced to 20-80 nm in typical dye. With this technique, material science and biomedical research are prosperously studied with nanometer resolution. The main work of this thesis is to build a STED microscopy based on a commercial confocal laser scanning microscope (CLSM) with the laser source of a supercontinuum. The resolution enhancement has been demonstrated by our home-built STED microscope. However, the resolution is confined by some aberrations existing in our system. The supercontinuum provides the capability in studying wide spectral range of fluorophore without the need of additional laser source. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T01:13:52Z (GMT). No. of bitstreams: 1 ntu-100-R98245009-1.pdf: 1441003 bytes, checksum: f9abae2e29822790a77c01b35fc1d43c (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | Thesis committee approvement
Acknowledgement i Chinese abstract ii English abstract iii Chapter 1 Introduction 7 1.1 Far-field Super-resolution Microscopy 7 1.1.1 Pointilism microscopy 7 1.1.2 STED Microscopy 8 1.2 Overview 8 Chapter 2 Theory 10 2.1 Diffraction Limit 10 2.2 Confocal Laser Scanning Microscope (CLSM) 12 2.3 Fluorescence 16 2.4 Stimulated Emssion 18 2.5 STED 19 2.5.1 Reversible Saturation Optical Fluorescence Transition (RESOLFT) 19 2.5.2 STED Microscopy 22 2.5.3 Beam Characteristic 23 2.5.3.1 Wavelength 23 2.5.3.2 Pulse width and Pulse Interval 24 2.5.3.3 Polarization 25 Chapter 3 Experiment Setup 26 3.1 Brief Introduction 26 3.2 Laser 27 3.3 Polarizing Cube 28 3.4 Select Wavelength 29 3.4.1 Wavelength Selector 29 3.4.2 Interference Filter 31 3.5 Fiber 32 3.6 Cage System 35 3.7 Vortex Phase Plate 36 3.8 Circular Polarization 37 3.9 Confocal Laser Scanning microscope (CLSM) 38 3.10 Time Correlated Single Photon Counting (TCSPC) 42 3.11 Sample Preparation 43 3.11.1 Gold Nanoparticle 43 3.11.2 Fluorescence sample 45 Chapter 4 Experiment Results 48 4.1 PSF Measurement and Spatial Overlap 48 4.2 Temporal Overlap 54 4.3 Depletion Test 56 4.3.1 Absorption of STED beam Test 56 4.3.1 On/Off Process 57 4.3.3 Depletion Curve 59 4.4 Resolution Test 62 Chapter 5 Discussion 68 Chapter 6 Conclusion 75 Figure Index 76 Table Index 78 Reference 79 | |
dc.language.iso | en | |
dc.title | 利用雷射掃描式共軛焦顯微鏡暨超連續雷射為光源
建構受激發射耗乏顯微鏡 | zh_TW |
dc.title | Construct a STED microscope on Confocal Laser Scanning Microscope With Supercontinuum Laser | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳永芳(Yang-Fang Chen),張煥正(Huan-Cheng Chang),劉子銘(Tzu-Ming Liu) | |
dc.subject.keyword | 受激發射耗乏,遠場光學顯微術,螢光顯微術,超解析,超連續雷射, | zh_TW |
dc.subject.keyword | STED,far-field optical microscopy,fluorescence microscopy,super-resolution,supercontinuum, | en |
dc.relation.page | 82 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-16 | |
dc.contributor.author-college | 理學院 | zh_TW |
dc.contributor.author-dept | 應用物理所 | zh_TW |
顯示於系所單位: | 應用物理研究所 |
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