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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: STORM成像突破了传统光学显微镜的衍射极限,能够以纳米级的精度观察亚细胞结构。在这一背景下,具有易于修饰的光响应二芳基乙烯类染料获得了新的发展机遇。然而目前用于STORM成像的二芳基乙烯类探针通常需要一束紫外光源调控荧光态与暗态之间的转变,而且缺少对细胞器具有定位效果的探针。本文提出了一种新颖的STORM荧光探针设计策略,将二芳基乙烯光响应单元与具有分子内质子转移效应的荧光团相结合,能够实现全可见光调控的荧光开关体系,在光响应荧光团另一侧修饰溶酶体的靶向单元,实现靶向溶酶体的全可见光调控STORM成像,单个溶酶体的STORM成像分辨率可达到99 nm。

       

      Abstract: Stochastic optical reconstruction microscopy (STORM) breaks through the diffraction limit of conventional optical imaging, allowing biological processes to be visualized with nanoscale precision. The diarylethene have attracted particular attention because of their remarkable reversibility, excellent thermal stability and fatigue resistance. Photoswitchable diarylethene with easy modification and tunable fluorescence eliminate the need of any additives in single-molecule localization techniques, and have gained new development opportunities in STORM imaging. However, most diarylethene probes currently used for STORM imaging usually require a beam of phototoxic ultraviolet (UV) laser to control the transition between the fluorescent and dark states, and also lack the capability of organelle recognition. Here, we propose a novel STORM fluorescence probe design strategy. By combing the intramolecular proton transfer (IPT) fluorophore on one side and the lysosomal targeting group on the other side with the diarylethene, we have successfully synthesized the molecule HMN which can not only regulate the fluorescence blinking between the bright and dark states by harmless all-visible light, but also have excellent lysosome targeting ability. These properties are benefit for the intracellular STORM imaging of lysosomes. HMN exhibits excellent fatigue resistance, sensitive fluorescence switching, accurate lysosome recognition capability, and its fluorescence can be activated by 488 nm laser and deactivated by using 561 nm laser without requiring any additives to the imaging media. With the photoresponsive probes HMN, we achieved STORM imaging with all-visible light, which contributes to determine the subcellular distribution of lysosomes and enable super-resolution of a single lysosome up to 99 nm.

       

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