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Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging

[Image: see text] Rhodamine dyes exist in equilibrium between a fluorescent zwitterion and a nonfluorescent lactone. Tuning this equilibrium toward the nonfluorescent lactone form can improve cell-permeability and allow creation of “fluorogenic” compounds—ligands that shift to the fluorescent zwitte...

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Autores principales: Zheng, Qinsi, Ayala, Anthony X., Chung, Inhee, Weigel, Aubrey V., Ranjan, Anand, Falco, Natalie, Grimm, Jonathan B., Tkachuk, Ariana N., Wu, Carl, Lippincott-Schwartz, Jennifer, Singer, Robert H., Lavis, Luke D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2019
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6764213/
https://www.ncbi.nlm.nih.gov/pubmed/31572787
http://dx.doi.org/10.1021/acscentsci.9b00676
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author Zheng, Qinsi
Ayala, Anthony X.
Chung, Inhee
Weigel, Aubrey V.
Ranjan, Anand
Falco, Natalie
Grimm, Jonathan B.
Tkachuk, Ariana N.
Wu, Carl
Lippincott-Schwartz, Jennifer
Singer, Robert H.
Lavis, Luke D.
author_facet Zheng, Qinsi
Ayala, Anthony X.
Chung, Inhee
Weigel, Aubrey V.
Ranjan, Anand
Falco, Natalie
Grimm, Jonathan B.
Tkachuk, Ariana N.
Wu, Carl
Lippincott-Schwartz, Jennifer
Singer, Robert H.
Lavis, Luke D.
author_sort Zheng, Qinsi
collection PubMed
description [Image: see text] Rhodamine dyes exist in equilibrium between a fluorescent zwitterion and a nonfluorescent lactone. Tuning this equilibrium toward the nonfluorescent lactone form can improve cell-permeability and allow creation of “fluorogenic” compounds—ligands that shift to the fluorescent zwitterion upon binding a biomolecular target. An archetype fluorogenic dye is the far-red tetramethyl-Si-rhodamine (SiR), which has been used to create exceptionally useful labels for advanced microscopy. Here, we develop a quantitative framework for the development of new fluorogenic dyes, determining that the lactone–zwitterion equilibrium constant (K(L–Z)) is sufficient to predict fluorogenicity. This rubric emerged from our analysis of known fluorophores and yielded new fluorescent and fluorogenic labels with improved performance in cellular imaging experiments. We then designed a novel fluorophore—Janelia Fluor 526 (JF(526))—with SiR-like properties but shorter fluorescence excitation and emission wavelengths. JF(526) is a versatile scaffold for fluorogenic probes including ligands for self-labeling tags, stains for endogenous structures, and spontaneously blinking labels for super-resolution immunofluorescence. JF(526) constitutes a new label for advanced microscopy experiments, and our quantitative framework will enable the rational design of other fluorogenic probes for bioimaging.
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spelling pubmed-67642132019-09-30 Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging Zheng, Qinsi Ayala, Anthony X. Chung, Inhee Weigel, Aubrey V. Ranjan, Anand Falco, Natalie Grimm, Jonathan B. Tkachuk, Ariana N. Wu, Carl Lippincott-Schwartz, Jennifer Singer, Robert H. Lavis, Luke D. ACS Cent Sci [Image: see text] Rhodamine dyes exist in equilibrium between a fluorescent zwitterion and a nonfluorescent lactone. Tuning this equilibrium toward the nonfluorescent lactone form can improve cell-permeability and allow creation of “fluorogenic” compounds—ligands that shift to the fluorescent zwitterion upon binding a biomolecular target. An archetype fluorogenic dye is the far-red tetramethyl-Si-rhodamine (SiR), which has been used to create exceptionally useful labels for advanced microscopy. Here, we develop a quantitative framework for the development of new fluorogenic dyes, determining that the lactone–zwitterion equilibrium constant (K(L–Z)) is sufficient to predict fluorogenicity. This rubric emerged from our analysis of known fluorophores and yielded new fluorescent and fluorogenic labels with improved performance in cellular imaging experiments. We then designed a novel fluorophore—Janelia Fluor 526 (JF(526))—with SiR-like properties but shorter fluorescence excitation and emission wavelengths. JF(526) is a versatile scaffold for fluorogenic probes including ligands for self-labeling tags, stains for endogenous structures, and spontaneously blinking labels for super-resolution immunofluorescence. JF(526) constitutes a new label for advanced microscopy experiments, and our quantitative framework will enable the rational design of other fluorogenic probes for bioimaging. American Chemical Society 2019-09-05 2019-09-25 /pmc/articles/PMC6764213/ /pubmed/31572787 http://dx.doi.org/10.1021/acscentsci.9b00676 Text en Copyright © 2019 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Zheng, Qinsi
Ayala, Anthony X.
Chung, Inhee
Weigel, Aubrey V.
Ranjan, Anand
Falco, Natalie
Grimm, Jonathan B.
Tkachuk, Ariana N.
Wu, Carl
Lippincott-Schwartz, Jennifer
Singer, Robert H.
Lavis, Luke D.
Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging
title Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging
title_full Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging
title_fullStr Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging
title_full_unstemmed Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging
title_short Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging
title_sort rational design of fluorogenic and spontaneously blinking labels for super-resolution imaging
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6764213/
https://www.ncbi.nlm.nih.gov/pubmed/31572787
http://dx.doi.org/10.1021/acscentsci.9b00676
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