Sparsity-based super-resolution microscopy from correlation information
Sparsity-based super-resolution microscopy from correlation information
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DOI：
10.1364/oe.26.018238
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发表时间：
2018-07-09
期刊：
影响因子：
3.8
通讯作者：
Eldar, Yonina C.
中科院分区：
文献类型：
--
作者：
Solomon, Oren;Mutzafi, Maor;Eldar, Yonina C.
一个多世纪以来，光的波长被认为是光学成像空间分辨率的基本限制。特别是在光学显微镜中，这个极限，被称为阿贝衍射极限，对高分辨率成像亚细胞细胞器的能力产生了根本性的限制。然而，现代显微镜技术，如STED， PALM和STORM，通过依赖荧光成像，设法恢复亚波长信息。具体来说，PALM/STORM从成像标本内附着在细胞器上的分子中获取大序列的荧光图像，这样在每帧中只有一小组荧光团是活跃的。每个荧光团的位置可以在每一帧中准确地找到，并通过叠加所有帧中的点来恢复图像。由此产生的颗粒状图像随后被平滑，以产生最终的分辨率为几十纳米的超分辨率图像。然而，由于PALM/STORM依赖于许多（大约10000）次曝光，它们的时间分辨率很差。为了解决这一问题，研究表明，通过允许更高的荧光团密度和利用发射的时间统计数据，超分辨率光学波动成像（SOFI）可以产生具有更高时间分辨率的亚衍射图像。但是，SOFI的时间分辨率提高是以牺牲空间分辨率为代价的，其空间分辨率不如PALM/STORM高。在这里，我们提出了一种新的方法，称为SPARCOM：基于稀疏性的超分辨率相关显微镜，它结合了比以前报道的更短的集成时间和与PALM和STORM相当的空间分辨率。SPARCOM依赖于相关域的稀疏性，利用荧光分子的稀疏分布和不同发射体之间缺乏相关性。我们在模拟和实验中展示了我们的技术，并提供了与最先进的高密度方法的比较。(C) 2018年美国光学学会根据OSA开放获取出版协议的条款
For more than a century, the wavelength of light was considered to be a fundamental limit on the spatial resolution of optical imaging. Particularly in light microscopy, this limit, known as Abbe's diffraction limit, places a fundamental constraint on the ability to image subcellular organelles with high resolution. However, modern microscopy techniques such as STED, PALM and STORM, manage to recover sub-wavelength information, by relying on fluorescence imaging. Specifically, PALM/STORM acquire large sequences of fluorescence images from molecules attached to the organelles within the imaged specimen, such that in each frame only a small set of fluorophores are active. The position of each fluorophore can be found accurately in each frame, and the image is recovered by superimposing the points from all frames. The resulting grainy image is subsequently smoothed to produce the final super-resolved image with a resolution of tens of nano-meters. However, because PALM/STORM rely on many (>10,000) exposures, they suffer from poor temporal resolution. To address that, super-resolution optical fluctuation imaging (SOFI) was shown to produce sub-diffraction images with increased temporal resolution, by allowing for higher fluorophore density and exploiting the temporal statistics of the emissions. However, the improved temporal resolution of SOFI comes at the expense of its spatial resolution, which is not as high as that of PALM/STORM. Here, we present a new method called SPARCOM: sparsity-based super-resolution correlation microscopy, which combines a shorter integration time than previously reported with spatial resolution comparable to PALM and STORM. SPARCOM relies on sparsity in the correlation domain, exploiting the sparse distribution of fluorescent molecules and the lack of correlation between different emitters. We demonstrate our technique in simulations and in experiments, and provide comparisons to state-of-the-art high density methods. (C) 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement