Phosphole-Oxide-Based Fluorescent Probe for Super-resolution Stimulated Emission Depletion Live Imaging of the Lysosome Membrane
Phosphole-Oxide-Based Fluorescent Probe for Super-resolution Stimulated Emission Depletion Live Imaging of the Lysosome Membrane
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DOI：
10.1021/acsmaterialslett.0c00147
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发表时间：
2020-07-06
影响因子：
11.4
通讯作者：
Yamaguchi, Shigehiro
中科院分区：
文献类型：
--
作者：
Wang, Chenguang;Taki, Masayasu;Yamaguchi, Shigehiro
超分辨率成像技术已成为越来越重要的工具，可视化亚细胞器结构和动态过程的活细胞在纳米尺度。然而，这些成像技术的应用目前受到先进荧光探针的限制，这些探针能够对感兴趣的细胞器进行特定标记，并提供超分辨率成像技术所需的吸收/发射特性。本文提出了一种新的小分子荧光探针LysoPB Yellow，它可以对溶酶体膜进行选择性染色。其优异的光稳定性和较长的荧光寿命有利于其在超分辨率受激发射耗尽（STED）显微镜中的应用。用STED成像技术在活细胞中成功地观察到用LysoPB Yellow染色的溶酶体膜，其全宽半宽（FWHM）为70 nm，基本低于光的衍射极限。此外，LysoPB Yellow具有良好的保留能力和可忽略的细胞毒性。因此，成功地进行了活细胞的长期延时共聚焦成像，证明了LysoPB Yellow在荧光成像中的实际应用。
Super-resolution imaging techniques have become increasingly important tools to visualize suborganelle structures and dynamic processes in living cells on the nanoscale. However, the utility of these imaging techniques is currently limited by the availability of advanced fluorescent probes that enable the specific labeling of the organelle of interest and provide the absorption/emission properties required for super-resolution imaging techniques. Herein, LysoPB Yellow is presented as a new small-molecule fluorescent probe that can selectivity stain the lysosomal membrane. Its outstandingly high photostability and long fluorescence lifetime are beneficial for its use in super-resolution stimulated emission depletion (STED) microscopy. A lysosomal membrane stained with LysoPB Yellow was successfully visualized by STED imaging in living cells with a full width at half maximum (FWHM) of 70 nm, which is substantially below the diffraction limit of light. Additionally, LysoPB Yellow displayed excellent retention ability and negligible cytotoxicity. Consequently, long-term time-lapse confocal imaging of living cells was successfully conducted, which demonstrates the practical utility of LysoPB Yellow in fluorescence imaging.