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Ultrabright photoactivatable fluorophores created by reductive caging

2012; Nature Portfolio; Volume: 9; Issue: 12 Linguagem: Inglês

10.1038/nmeth.2214

1548-7105

Joshua C. Vaughan, Shu Jia, Xiaowei Zhuang,

Advanced Electron Microscopy Techniques and Applications

The resolution achievable with single-molecule–based super-resolution fluorescence imaging is increased via a fluorophore caging procedure that uses a reducing agent to convert dyes to a long-lived dark state from which they can be activated with UV light and emit high numbers of photons. Sub–diffraction-limit imaging can be achieved by sequential localization of photoactivatable fluorophores, for which the image resolution depends on the number of photons detected per localization. We report a strategy for fluorophore caging that creates photoactivatable probes with high photon yields. Upon photoactivation, these probes can provide 104−106 photons per localization and allow imaging of fixed samples with resolutions of several nanometers. This strategy can be applied to many fluorophores across the visible spectrum.