Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2011-12

AUTHORS

Graham T Dempsey, Joshua C Vaughan, Kok Hao Chen, Mark Bates, Xiaowei Zhuang

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. More... »

PAGES

1027

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nmeth.1768

DOI

http://dx.doi.org/10.1038/nmeth.1768

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1043491552

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/22056676


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