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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2011
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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. |
format | Online Article Text |
id | pubmed-3272503 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
record_format | MEDLINE/PubMed |
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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