Spontaneously blinking fluorophores based on nucleophilic addition/dissociation of intracellular glutathione for live-cell super-resolution imaging,Journal of the American Chemical Society

当前位置： X-MOL 学术 › J. Am. Chem. Soc. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Spontaneously blinking fluorophores based on nucleophilic addition/dissociation of intracellular glutathione for live-cell super-resolution imaging
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-05-12 , DOI: 10.1021/jacs.0c00451
Akihiko Morozumi , Mako Kamiya ₁ , Shin-Nosuke Uno , Keitaro Umezawa , Ryosuke Kojima ₁ , Toshitada Yoshihara ₂ , Seiji Tobita ₂ , Yasuteru Urano ₃

Affiliation

Single-molecule localization microscopy (SMLM) allows the reconstruction of super-resolution images, but generally requires prior intense laser irradiation and in some cases additives to induce blinking of conventional fluorophores. We previously introduced a spontaneously blinking rhodamine fluorophore based on an intramolecular spirocyclization reaction for live-cell SMLM under physiological conditions. Here, we report a novel principle of spontaneous blinking in living cells, which utilizes reversible ground-state nucleophilic attack of intracellular glutathione (GSH) upon a xanthene fluorophore. Structural optimization afforded two pyronine fluorophores with different colors, both of which exhibit equilibrium (between the fluorescent dissociated form and the non-fluorescent GSH adduct form) and blinking kinetics that enable SMLM of microtubules in living cells. Furthermore, by using spontaneously blinking fluorophores working in the near-infrared (NIR) and green ranges, we succeeded in dual-color live-cell SMLM without the need for optimization of the imaging medium.

中文翻译：

基于细胞内谷胱甘肽亲核添加/解离的自发闪烁荧光团用于活细胞超分辨率成像

单分子定位显微镜 (SMLM) 允许重建超分辨率图像，但通常需要事先进行强激光照射，在某些情况下还需要添加剂来诱导常规荧光团的闪烁。我们之前基于活细胞 SMLM 在生理条件下的分子内螺环化反应引入了自发闪烁的罗丹明荧光团。在这里，我们报告了活细胞中自发闪烁的新原理，该原理利用细胞内谷胱甘肽 (GSH) 对呫吨荧光团的可逆基态亲核攻击。结构优化提供了两种不同颜色的吡罗宁荧光团，两者都表现出平衡（在荧光解离形式和非荧光 GSH 加合物形式之间）和闪烁动力学，使活细胞中的微管能够进行 SMLM。此外，通过使用在近红外 (NIR) 和绿色范围内工作的自发闪烁荧光团，我们成功地实现了双色活细胞 SMLM，而无需优化成像介质。

更新日期：2020-05-12

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南