Symmetrically dispersed spectroscopic single-molecule localization microscopy
Symmetrically dispersed spectroscopic single-molecule localization microscopy
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DOI：
10.1038/s41377-020-0333-9
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发表时间：
2020-05-25
影响因子：
19.4
通讯作者：
Zhang, Hao F.
中科院分区：
文献类型：
--
作者：
Song, Ki-Hee;Zhang, Yang;Zhang, Hao F.
光谱单分子定位显微镜（sSMLM）首次实现了单分子的同时成像和光谱分析。目前的sSMLM从根本上遭受减少的光子预算，因为来自个体随机发射的光子被分成空间和光谱通道。因此，空间定位和光谱分析都只使用总光子的一部分，导致两个通道的精度降低。为了提高空间和光谱精度，我们提出了对称分散的sSMLM，或SDsSMLM，充分利用所有的光子从个人的随机发射在空间和光谱通道。SDsSMLM实现了10纳米的空间和0.8纳米的光谱精度在总光子预算为1000。与现有的sSMLM使用1：3的空间和光谱通道之间的分裂比相比，SDsSMLM提高了空间和光谱精度分别为42%和10%，在相同的光子预算。我们还展示了固定细胞的多色成像和使用SDsSMLM的三维单粒子跟踪。SDsSMLM可在更广泛的细胞生物学和材料科学应用中实现更精确的光谱单分子分析。显微镜：改进单分子定位和分析研究人员改进了称为光谱单分子定位显微镜（sSMLM）的技术，可以同时定位单个分子，并获得携带分子结构信息的光谱特征。该方法由Hao F.美国伊利诺斯州西北大学的Zhang及其同事是第一个使用分子发射的所有光子来提供空间和光谱信息的人。以前，光子在单独的空间和光谱通道中传递的信息不能组合使用。这项技术被称为对称分散sSMLM，空间精度提高了42%，光谱精度提高了10%。研究人员还展示了细胞中多色成像和三维跟踪监测纳米颗粒的技术。它将显著增强生物学和材料科学中单分子水平的光谱分析。
Spectroscopic single-molecule localization microscopy (sSMLM) was used to achieve simultaneous imaging and spectral analysis of single molecules for the first time. Current sSMLM fundamentally suffers from a reduced photon budget because the photons from individual stochastic emissions are divided into spatial and spectral channels. Therefore, both spatial localization and spectral analysis only use a portion of the total photons, leading to reduced precisions in both channels. To improve the spatial and spectral precisions, we present symmetrically dispersed sSMLM, or SDsSMLM, to fully utilize all photons from individual stochastic emissions in both spatial and spectral channels. SDsSMLM achieved 10-nm spatial and 0.8-nm spectral precisions at a total photon budget of 1000. Compared with the existing sSMLM using a 1:3 splitting ratio between spatial and spectral channels, SDsSMLM improved the spatial and spectral precisions by 42% and 10%, respectively, under the same photon budget. We also demonstrated multicolour imaging of fixed cells and three-dimensional single-particle tracking using SDsSMLM. SDsSMLM enables more precise spectroscopic single-molecule analysis in broader cell biology and material science applications.Microscopy: Improved localization and analysis of single molecules Researchers have refined the technique called spectroscopic single-molecule localization microscopy (sSMLM) to simultaneously locate individual molecules and also obtain spectral signatures carrying information about their molecular structures. The method, developed by Hao F. Zhang and colleagues at Northwestern University in Illinois, USA, is the first to use of all of the photons emitted by molecules to provide both spatial and spectral information. Previously the information delivered by photons in separate spatial and spectral channels could not be used in combination. The technique, called Symmetrically dispersed sSMLM, achieves a 42 percent increase in spatial precision and a 10 percent increase in spectral precision. The researchers also demonstrated the technique for multicolour imaging in cells and three-dimensional tracking for monitoring nanoparticles. It should significantly enhance spectroscopic analysis at the single-molecule level in both biology and materials science.