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Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe

Recent developments in fluorescence microscopy call for novel small‐molecule‐based labels with multiple functionalities to satisfy different experimental requirements. A current limitation in the advancement of live‐cell single‐molecule localization microscopy is the high excitation power required t...

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Autores principales: Werther, Philipp, Yserentant, Klaus, Braun, Felix, Kaltwasser, Nicolai, Popp, Christoph, Baalmann, Mathis, Herten, Dirk‐Peter, Wombacher, Richard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6972563/
https://www.ncbi.nlm.nih.gov/pubmed/31638314
http://dx.doi.org/10.1002/anie.201906806
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author Werther, Philipp
Yserentant, Klaus
Braun, Felix
Kaltwasser, Nicolai
Popp, Christoph
Baalmann, Mathis
Herten, Dirk‐Peter
Wombacher, Richard
author_facet Werther, Philipp
Yserentant, Klaus
Braun, Felix
Kaltwasser, Nicolai
Popp, Christoph
Baalmann, Mathis
Herten, Dirk‐Peter
Wombacher, Richard
author_sort Werther, Philipp
collection PubMed
description Recent developments in fluorescence microscopy call for novel small‐molecule‐based labels with multiple functionalities to satisfy different experimental requirements. A current limitation in the advancement of live‐cell single‐molecule localization microscopy is the high excitation power required to induce blinking. This is in marked contrast to the minimal phototoxicity required in live‐cell experiments. At the same time, quality of super‐resolution imaging depends on high label specificity, making removal of excess dye essential. Approaching both hurdles, we present the design and synthesis of a small‐molecule label comprising both fluorogenic and self‐blinking features. Bioorthogonal click chemistry ensures fast and highly selective attachment onto a variety of biomolecular targets. Along with spectroscopic characterization, we demonstrate that the probe improves quality and conditions for regular and single‐molecule localization microscopy on live‐cell samples.
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spelling pubmed-69725632020-01-27 Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe Werther, Philipp Yserentant, Klaus Braun, Felix Kaltwasser, Nicolai Popp, Christoph Baalmann, Mathis Herten, Dirk‐Peter Wombacher, Richard Angew Chem Int Ed Engl Research Articles Recent developments in fluorescence microscopy call for novel small‐molecule‐based labels with multiple functionalities to satisfy different experimental requirements. A current limitation in the advancement of live‐cell single‐molecule localization microscopy is the high excitation power required to induce blinking. This is in marked contrast to the minimal phototoxicity required in live‐cell experiments. At the same time, quality of super‐resolution imaging depends on high label specificity, making removal of excess dye essential. Approaching both hurdles, we present the design and synthesis of a small‐molecule label comprising both fluorogenic and self‐blinking features. Bioorthogonal click chemistry ensures fast and highly selective attachment onto a variety of biomolecular targets. Along with spectroscopic characterization, we demonstrate that the probe improves quality and conditions for regular and single‐molecule localization microscopy on live‐cell samples. John Wiley and Sons Inc. 2019-11-18 2020-01-07 /pmc/articles/PMC6972563/ /pubmed/31638314 http://dx.doi.org/10.1002/anie.201906806 Text en © 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Werther, Philipp
Yserentant, Klaus
Braun, Felix
Kaltwasser, Nicolai
Popp, Christoph
Baalmann, Mathis
Herten, Dirk‐Peter
Wombacher, Richard
Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe
title Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe
title_full Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe
title_fullStr Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe
title_full_unstemmed Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe
title_short Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe
title_sort live‐cell localization microscopy with a fluorogenic and self‐blinking tetrazine probe
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6972563/
https://www.ncbi.nlm.nih.gov/pubmed/31638314
http://dx.doi.org/10.1002/anie.201906806
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