Fast stimulated emission nanoscopy based on single molecule localization.
Fast stimulated emission nanoscopy based on single molecule localization.
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DOI：
10.1364/ao.54.006919
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发表时间：
2015-08
期刊：
影响因子：
1.9
通讯作者：
Xuehua Wang;Danni Chen;Bin Yu;H. Niu
中科院分区：
文献类型：
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作者：
Xuehua Wang;Danni Chen;Bin Yu;H. Niu
对于基于单分子随机开关和局域化的超分辨显微镜方法，为了同时提高时空分辨率，必须最大化单位时间内单分子可以收集的光子数。在这里，我们描述了一种新的方法，通过引入受激发射(SE)光学过程来增强荧光探针的信号强度(每秒收集的光子数)。这一过程基于以下两个性质：第一，在合理的参数下，SE可以显著提高光子发射率；第二，SE光子在空间上与刺激光束相干，比荧光更有利于收集。理论结果表明，SE可以大大提高单个荧光分子的信号强度。因此，我们表明，结合单分子定位方法，使用SE可以获得每个重建图像0.05S的快速成像，横向分辨率为∼30 nm。
For super-resolution microscopy methods based on single molecule stochastic switching and localization, to simultaneously improve the spatial-temporal resolution, it is necessary to maximize the number of photons that can be collected from single molecules per unit time. Here, we describe a novel approach to enhance the signal intensity (collected photons per second) from fluorescence probes by introducing a stimulated emission (SE) optical process. This process is based on the following two properties: first, with reasonable parameters, the photon emission rate can be significantly increased with SE; and second, the SE photons, which are spatially coherent with the stimulation beam, are more favorable for collection than fluorescence. Theoretical results have shown that signal intensity from a single fluorescent molecule can be greatly improved with SE. We therefore showed, using SE in combination with single molecule localization methodology, that fast imaging at a rate of 0.05 s per reconstructed image with lateral resolutions of ∼30 nm can be obtained.