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<p>Stimulated emission depletion (STED) microscopy, a key optical super-resolution imaging method, has extended our ability to view details to resolution levels of tens of nanometers. Its resolution depends on fluorophore de-excitation efficiency, and increases with depletion laser power. However, high-power irradiation permanently turns off the fluorescence due to photo-bleaching of the fluorophores. As a result, there is a trade-off between spatial resolution and imaging time. Here, we overcome this limitation by introducing reactivatable STED (ReSTED) based on the photophysical properties of the nanographene dibenzo[<i>hi</i>,<i>st</i>]ovalene (DBOV). In contrast to the photo-induced decomposition of other fluorophores, the fluorescence of DBOV is only temporarily deactivated and can be reactivated by near-infrared light (including the 775 nm depletion beam). As a result, this fluorophore allows for hours-long, high-resolution 3D STED imaging, greatly expanding the applications of STED microscopy.</p>

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Reactivatable stimulated emission depletion microscopy using fluorescence-recoverable nanographene

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年2月4日

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  • Qiqi Yang,
  • Antonio Virgilio Failla,
  • Petri Turunen,
  • Ana Mateos-Maroto,
  • Meiyu Gai,
  • Werner Zuschratter,
  • Sophia Westendorf,
  • Márton Gelléri,
  • Qiang Chen,
  • Goudappagouda,
  • Hao Zhao,
  • Xingfu Zhu,
  • Svenja Morsbach,
  • Marcus Scheele,
  • Wei Yan,
  • Katharina Landfester,
  • Ryota Kabe,
  • Mischa Bonn,
  • Akimitsu Narita,
  • Xiaomin Liu

摘要

Stimulated emission depletion (STED) microscopy, a key optical super-resolution imaging method, has extended our ability to view details to resolution levels of tens of nanometers. Its resolution depends on fluorophore de-excitation efficiency, and increases with depletion laser power. However, high-power irradiation permanently turns off the fluorescence due to photo-bleaching of the fluorophores. As a result, there is a trade-off between spatial resolution and imaging time. Here, we overcome this limitation by introducing reactivatable STED (ReSTED) based on the photophysical properties of the nanographene dibenzo[hi,st]ovalene (DBOV). In contrast to the photo-induced decomposition of other fluorophores, the fluorescence of DBOV is only temporarily deactivated and can be reactivated by near-infrared light (including the 775 nm depletion beam). As a result, this fluorophore allows for hours-long, high-resolution 3D STED imaging, greatly expanding the applications of STED microscopy.