Subdiffraction resolution in far-field fluorescence microscopy
Subdiffraction resolution in far-field fluorescence microscopy
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DOI：
10.1364/ol.24.000954
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发表时间：
1999-07-15
期刊：
影响因子：
3.6
通讯作者：
Hell, SW
中科院分区：
文献类型：
--
作者：
Klar, TA;Hell, SW
我们克服了扫描远场荧光显微镜的分辨率限制，使焦斑外部的荧光失效。近紫外脉冲产生受激分子的衍射限制分布，而空间偏移脉冲通过受激发射从焦点的外部淬灭受激分子。这就产生了子衍射大小的有效点扩展函数。对于1.4孔径和388 nm激发波长，单偏移光束的空间分辨率从150 +/- 8 nm增加到106 +/- 8 nm。优异的横向分辨率被证明是通过分离相邻的吡啶2纳米晶体，否则是无法分辨的。(C) 1999美国光学学会OCIS代码：000.3110,180.2520,350.5730,320.7150。
We overcame the resolution limit of scanning far-field fluorescence microscopy by disabling the fluorescence from the outer part of the focal spot. Whereas a near-UV pulse generates a diffraction-limited distribution of excited molecules, a spatially offset pulse quenches the excited molecules from the outer part of the focus through stimulated emission. This results in a subdiffraction-sized effective point-spread function. For a 1.4 aperture and a 388-nm excitation wavelength spatial resolution is increased from 150 +/- 8 nm to 106 +/- 8 nm with a single offset beam. Superior lateral resolution is demonstrated by separation of adjacent Pyridine 2 nanocrystals that are otherwise indiscernible. (C) 1999 Optical Society of America OCIS codes: 000.3110, 180.2520, 350.5730, 320.7150.