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

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/o...

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Detalles Bibliográficos
Autores principales: Dempsey, Graham T., Vaughan, Joshua C., Chen, Kok Hao, Bates, Mark, Zhuang, Xiaowei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3272503/
https://www.ncbi.nlm.nih.gov/pubmed/22056676
http://dx.doi.org/10.1038/nmeth.1768
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author Dempsey, Graham T.
Vaughan, Joshua C.
Chen, Kok Hao
Bates, Mark
Zhuang, Xiaowei
author_facet Dempsey, Graham T.
Vaughan, Joshua C.
Chen, Kok Hao
Bates, Mark
Zhuang, Xiaowei
author_sort Dempsey, Graham T.
collection PubMed
description 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. While 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 a set of 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 crosstalk, four-color super-resolution imaging.
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spelling pubmed-32725032012-06-01 Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging Dempsey, Graham T. Vaughan, Joshua C. Chen, Kok Hao Bates, Mark Zhuang, Xiaowei Nat Methods Article 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. While 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 a set of 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 crosstalk, four-color super-resolution imaging. 2011-11-06 /pmc/articles/PMC3272503/ /pubmed/22056676 http://dx.doi.org/10.1038/nmeth.1768 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Dempsey, Graham T.
Vaughan, Joshua C.
Chen, Kok Hao
Bates, Mark
Zhuang, Xiaowei
Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
title Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
title_full Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
title_fullStr Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
title_full_unstemmed Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
title_short Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
title_sort evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3272503/
https://www.ncbi.nlm.nih.gov/pubmed/22056676
http://dx.doi.org/10.1038/nmeth.1768
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