化学
荧光团
生物物理学
谷胱甘肽
细胞内
荧光
亲核细胞
光化学
活体细胞成像
罗丹明
杂蒽
细胞
生物化学
光学
催化作用
酶
物理
生物
作者
Akihiko Morozumi,Mako Kamiya,Shin‐nosuke Uno,Keitaro Umezawa,Ryosuke Kojima,Toshitada Yoshihara,Seiji Tobita,Yasuteru Urano
摘要
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 nonfluorescent GSH adduct form) and blinking kinetics that enable SMLM of microtubules or mitochondria 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.
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