Li Fanghui, Li Mengqi, Shi Yiqi, Bian Xinyun, Lv Ning, Guo Shaomeng, Wang Ying, Zhao Weijun, Zhu Wei-Hong
Key Laboratory for Advanced Materials and Joint International Research Laboratory of Precision Chemistry and Molecular Engineering, Feringa Nobel Prize Scientist Joint Research Center, Shanghai Key Laboratory of Functional Materials Chemistry, Institute of Fine Chemicals, East China University of Science & Technology Shanghai 200237 China
Center of Photosensitive Chemicals Engineering, East China University of Science and Technology Shanghai 200237 China.
Chem Sci. 2025 Jul 10. doi: 10.1039/d5sc03224e.
Stochastic optical reconstruction microscopy (STORM) overcomes the diffraction limit of optical imaging, facilitating high-resolution visualization of cellular substructures at the nanoscale. Essential to this technique is the development of fluorescent photoswitches. However, existing photoswitches typically rely on sophisticated dual-beam systems that involve harmful UV-light and lack specific recognition of biomolecules. Here we develop unique intracellular biomolecule-activatable photoswitches tailored for single-visible-light performed STORM imaging. Upon incorporating intramolecular proton transfer (IPT) units into the photochromic diarylethene, the all-visible-light driven photoswitches are established with excellent photoresponsive efficiency, high brightness and fluorescence ON-to-OFF contrast ratio, guaranteeing STORM imaging using a single-visible-light (488 nm) by regulating the activation, excitation and deactivation processes. Furthermore, we functionalized the IPT units with biomolecular recognition motifs, creating photoswitches capable of sensing the expression levels of intracellular biomolecules (like glutathione (GSH) or β-galactosidase (β-Gal)) with super-resolution. Our objective is to engineer single-visible-light driven, biomolecule-activatable photoswitches, which will significantly streamline the STORM technique and expand the applicability of super-resolution imaging for the precise mapping of intracellular substructures.
随机光学重建显微镜（STORM）克服了光学成像的衍射极限，有助于在纳米尺度上对细胞亚结构进行高分辨率可视化。该技术的关键在于荧光光开关的开发。然而，现有的光开关通常依赖于复杂的双光束系统，该系统涉及有害的紫外线，并且缺乏对生物分子的特异性识别。在此，我们开发了独特的可被细胞内生物分子激活的光开关，专为单可见光进行的STORM成像量身定制。通过将分子内质子转移（IPT）单元引入到光致变色二芳基乙烯中，建立了全可见光驱动的光开关，其具有优异的光响应效率、高亮度和荧光开-关对比度，通过调节激活、激发和失活过程，保证了使用单可见光（488nm）进行STORM成像。此外，我们用生物分子识别基序对IPT单元进行功能化，创造出能够以超分辨率检测细胞内生物分子（如谷胱甘肽（GSH）或β-半乳糖苷酶（β-Gal））表达水平的光开关。我们的目标是设计出单可见光驱动、可被生物分子激活的光开关，这将显著简化STORM技术，并扩大超分辨率成像在细胞内亚结构精确映射方面的适用性。
