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

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。

關鍵字

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

並列摘要

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.

並列關鍵字

STED ； far-field optical microscopy ； fluorescence microscopy ； super-resolution ； supercontinuum

參考文獻

1. S. W. Hell, "Far-field Optical Nanoscopy", Science 316, 1153 (2007).

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).

國際替代計量

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

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