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.

中文翻译：

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基于磷氧化物的荧光探针用于溶酶体膜的超高分辨率刺激发射耗尽实时成像

超分辨率成像技术已成为越来越重要的工具，可以在纳米尺度上可视化活细胞中的亚细胞器结构和动态过程。然而，这些成像技术的实用性当前受到先进荧光探针的可用性的限制，所述先进荧光探针能够对感兴趣的细胞器进行特异性标记并提供超分辨率成像技术所需的吸收/发射特性。在本文中，LysoPB Yellow被提出作为一种新型的小分子荧光探针，可以选择性地染色溶酶体膜。其出色的高光稳定性和长荧光寿命有利于将其用于超分辨率受激发射损耗（STED）显微镜。通过STED成像，在活细胞中成功地观察到了用LysoPB Yellow染色的溶酶体膜，其半高全宽（FWHM）为70 nm，该宽度基本上低于光的衍射极限。另外，LysoPB Yellow显示出优异的保留能力和可忽略的细胞毒性。因此，成功进行了活细胞的长期延时共聚焦成像，证明了LysoPB Yellow在荧光成像中的实用性。