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    全可见光调控二芳基乙烯光响应探针用于溶酶体STORM成像

    All-Visible-Light Activated Diarylethene Photoresponsive Probe for Lysosomal STORM Imaging

    • 摘要: 随机光学重建显微镜(STORM)突破了传统光学成像的衍射极限,可实现纳米级精度的生物成像。二芳基乙烯因其可逆性、热稳定性和抗疲劳性而成为重要的光开关材料,但现有 STORM 探针多依赖光毒性的紫外光调控荧光态,且缺乏细胞器靶向能力。为解决这些问题,本文提出一种新型 STORM 探针设计策略:在二芳基乙烯一侧引入具有分子内质子转移(IPT)效应的荧光团,另一侧修饰溶酶体靶向基团,成功构建了全可见光调控的探针 HMN。HMN 可在 488 nm 激光下激活荧光,在 561 nm 光照下关闭荧光,实现无需添加剂的明暗态可逆切换,并表现出优异的抗疲劳性和灵敏的光开关特性。同时,HMN 具有准确的溶酶体靶向能力,可用于活细胞中溶酶体的高分辨 STORM 成像。基于 HMN 的全可见光 STORM 成像不仅能够描绘溶酶体的亚细胞分布,而且实现了单个溶酶体高达 99 nm 的成像分辨率。

       

      Abstract: Stochastic optical reconstruction microscopy (STORM) breaks through the diffraction limit of conventional optical imaging, enabling visualization of biological processes with nanoscale precision. Diarylethene derivatives have attracted particular attention due to their remarkable reversibility, excellent thermal stability, and fatigue resistance. Photoswitchable diarylethene compounds with easy modification and tunable fluorescence eliminate the need for additives in single-molecule localization techniques, opening new development opportunities in STORM imaging. However, most diarylethene probes currently used for STORM imaging typically require a phototoxic ultraviolet (UV) laser to regulate the transition between fluorescent and dark states, and they lack organelle recognition capability. Herein, we propose a novel design strategy for STORM fluorescent probes. By conjugating an intramolecular proton transfer (IPT) fluorophore on one side and a lysosomal targeting group on the other with diarylethene, we successfully synthesized the molecule HMN. This probe not only regulates fluorescence blinking between bright and dark states using harmless all-visible light but also exhibits excellent lysosome-targeting capability, which facilitates intracellular STORM imaging of lysosomes. HMN demonstrates outstanding fatigue resistance, sensitive fluorescence switching, and accurate lysosome recognition. Its fluorescence can be activated by a 488 nm laser and deactivated by a 561 nm laser, without requiring any additives in the imaging medium. Using the photoresponsive probe HMN, we achieved all-visible-light STORM imaging, which helps determine the subcellular distribution of lysosomes and enables super-resolution imaging of individual lysosomes with a resolution of up to 99 nm.

       

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