Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion

Zhang Yang; Song Ki-Hee; Dong Biqin; Davis Janel L.; Shao Guangbin; Sun Cheng; Zhang Hao F.

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Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion

机译：多色超分辨率成像使用光谱单分子定位显微镜具有最佳光谱分散体

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We developed transmission diffraction grating-based spectroscopic single-molecule localization microscopy (sSMLM) to collect the spatial and spectral information of single-molecule blinking events concurrently. We characterized the spectral heterogeneities of multiple far-red emitting dyes in a high-throughput manner using sSMLM. We also investigated the influence of spectral dispersion on the single-molecule identification performance of fluorophores with large spectral overlapping. The careful tuning of spectral dispersion in grating-based sSMLM permitted simultaneous three-color super-resolution imaging in fixed cells with a single objective lens at a relatively low photon budget. Our sSMLM has a compact optical design and can be integrated with conventional localization microscopy to provide add-on spectroscopic analysis capability. (C) 2019 Optical Society of America

机译：我们开发了传输衍射光栅的光谱单分子定位显微镜（SSMLM），以同时收集单分子闪烁事件的空间和光谱信息。我们使用SSMLM以高吞吐量方式表征多个远红发射染料的光谱异质性。我们还研究了光谱分子对具有大光谱重叠的荧光团的单分子鉴定性能的影响。在光栅的SSMLM中仔细调整光谱色散在相对低的光子预算中具有单个物镜的固定电池中的同时三色超分辨率成像。我们的SSMLM具有紧凑的光学设计，可以与常规定位显微镜集成，以提供附加光谱分析能力。（c）2019年光学学会

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来源
《Applied optics》 |2019年第9期|共8页
作者
Zhang Yang; Song Ki-Hee; Dong Biqin; Davis Janel L.; Shao Guangbin; Sun Cheng; Zhang Hao F.;
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Northwestern Univ Dept Biomed Engn 2145 Sheridan Rd Evanston IL 60208 USA;

Northwestern Univ Dept Biomed Engn 2145 Sheridan Rd Evanston IL 60208 USA;

Northwestern Univ Dept Biomed Engn 2145 Sheridan Rd Evanston IL 60208 USA;

Northwestern Univ Dept Biomed Engn 2145 Sheridan Rd Evanston IL 60208 USA;

Northwestern Univ Dept Mech Engn 2145 Sheridan Rd Evanston IL 60208 USA;

Northwestern Univ Dept Mech Engn 2145 Sheridan Rd Evanston IL 60208 USA;

Northwestern Univ Dept Biomed Engn 2145 Sheridan Rd Evanston IL 60208 USA;

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Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion

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