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Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging.


ABSTRACT: One approach to super-resolution fluorescence imaging uses sequential activation and localization of individual fluorophores to achieve high spatial resolution. Essential to this technique is the choice of fluorescent probes; the properties of the probes, including photons per switching event, on-off duty cycle, photostability and number of switching cycles, largely dictate the quality of super-resolution images. Although many probes have been reported, a systematic characterization of the properties of these probes and their impact on super-resolution image quality has been described in only a few cases. Here we quantitatively characterized the switching properties of 26 organic dyes and directly related these properties to the quality of super-resolution images. This analysis provides guidelines for characterization of super-resolution probes and a resource for selecting probes based on performance. Our evaluation identified several photoswitchable dyes with good to excellent performance in four independent spectral ranges, with which we demonstrated low-cross-talk, four-color super-resolution imaging.

SUBMITTER: Dempsey GT 

PROVIDER: S-EPMC3272503 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature

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Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging.

Dempsey Graham T GT   Vaughan Joshua C JC   Chen Kok Hao KH   Bates Mark M   Zhuang Xiaowei X  

Nature methods 20111106 12


One approach to super-resolution fluorescence imaging uses sequential activation and localization of individual fluorophores to achieve high spatial resolution. Essential to this technique is the choice of fluorescent probes; the properties of the probes, including photons per switching event, on-off duty cycle, photostability and number of switching cycles, largely dictate the quality of super-resolution images. Although many probes have been reported, a systematic characterization of the prope  ...[more]

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