Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI)
Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI)
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DOI：
10.1073/pnas.0907866106
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发表时间：
2009-12-29
影响因子：
11.1
通讯作者：
Enderlein, J.
中科院分区：
文献类型：
--
作者：
Dertinger, T.;Colyer, R.;Enderlein, J.
超分辨光学显微镜是荧光显微镜领域中一个快速发展的领域，在影响众多科学领域方面具有巨大潜力。在过去十年中已经开发了几种超分辨方法，它们都能够克服光的基本衍射极限。在此我们提出一种在所有三个维度上获得亚衍射极限光学分辨率的方法。这种方法依赖于对一系列图像（影片）中记录的时间波动（由荧光闪烁/间歇性引起）进行高阶统计分析。我们通过使用常规的宽场显微镜证明了空间分辨率提高了5倍。这种分辨率的提高是通过迭代离散步骤实现的，这反过来又允许对不同分辨率水平的图像进行评估。即使在分辨率提高的最低水平，我们的方法也具有显著的背景降低以及因此而产生的对比度增强的特点，并且在成纤维细胞的量子点标记微管上得到了验证。
Super-resolution optical microscopy is a rapidly evolving area of fluorescence microscopy with a tremendous potential for impacting many fields of science. Several super-resolution methods have been developed over the last decade, all capable of overcoming the fundamental diffraction limit of light. We present here an approach for obtaining subdiffraction limit optical resolution in all three dimensions. This method relies on higher-order statistical analysis of temporal fluctuations (caused by fluorescence blinking/intermittency) recorded in a sequence of images (movie). We demonstrate a 5-fold improvement in spatial resolution by using a conventional wide-field microscope. This resolution enhancement is achieved in iterative discrete steps, which in turn allows the evaluation of images at different resolution levels. Even at the lowest level of resolution enhancement, our method features significant background reduction and thus contrast enhancement and is demonstrated on quantum dot-labeled microtubules of fibroblast cells.