A synergistic strategy to develop photostable and bright dyes with long Stokes shift for nanoscopy.

A synergistic strategy to develop photostable and bright dyes with long Stokes shift for nanoscopy.
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DOI:
10.1038/s41467-022-29547-3
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发表时间:
2022-04-27
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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--
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超分辨率荧光成像的质量和应用很大程度上取决于染料的性能,包括光稳定性、亮度和斯托克斯位移。在这里,我们报告了一种协同策略,可以同时改善常规荧光团的此类特性。将具有微调电子密度的喹喔啉基序引入到传统罗丹明中,生成具有振动结构的新型染料,并同时抑制扭曲分子内电荷转移(TICT)的形成,从而在增加斯托克斯位移的同时提高亮度和光稳定性。新型荧光团 YL578 的亮度和斯托克斯位移比其亲代荧光团罗丹明 B 大约高出两倍。重要的是,在受激发射损耗 (STED) 显微镜中,YL578 衍生的探针具有卓越的光稳定性,因此比基于卡吡喃宁的探针(CPY-Halo 和 580CP-Halo)(被称为 STED 成像的光稳定荧光团)呈现多三倍的帧数。此外,该策略得到了很好的推广,为生物成像和生物传感提供了一种具有长斯托克斯位移(高达 136 nm)的新型明亮且光稳定的荧光探针。超分辨率显微镜是细胞研究的强大工具,但需要明亮且稳定的荧光探针。在这里,作者报告了一种将喹喔啉基序引入传统探针的策略,使它们更亮、更耐光、更大的斯托克斯位移,并展示了用于生物传感应用的探针。
The quality and application of super-resolution fluorescence imaging greatly lie in the dyes’ properties, including photostability, brightness, and Stokes shift. Here we report a synergistic strategy to simultaneously improve such properties of regular fluorophores. Introduction of quinoxaline motif with fine-tuned electron density to conventional rhodamines generates new dyes with vibration structure and inhibited twisted-intramolecular-charge-transfer (TICT) formation synchronously, thus increasing the brightness and photostability while enlarging Stokes shift. The new fluorophore YL578 exhibits around twofold greater brightness and Stokes shift than its parental fluorophore, Rhodamine B. Importantly, in Stimulated Emission Depletion (STED) microscopy, YL578 derived probe possesses a superior photostability and thus renders threefold more frames than carbopyronine based probes (CPY-Halo and 580CP-Halo), known as photostable fluorophores for STED imaging. Furthermore, the strategy is well generalized to offer a new class of bright and photostable fluorescent probes with long Stokes shift (up to 136 nm) for bioimaging and biosensing. Super-resolution microscopy is a powerful tool for cellular studies but requires bright and stable fluorescent probes. Here, the authors report on a strategy to introduce quinoxaline motifs to conventional probes to make them brighter, more photostable, larger Stokes shift, and demonstrate the probes for biosensing applications.
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