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Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy
[Image: see text] Small-molecule fluorophores enable the observation of biomolecules in their native context with fluorescence microscopy. Specific labeling via bio-orthogonal tetrazine chemistry combines minimal label size with rapid labeling kinetics. At the same time, fluorogenic tetrazine–dye co...
Autores principales: | , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8461766/ https://www.ncbi.nlm.nih.gov/pubmed/34584958 http://dx.doi.org/10.1021/acscentsci.1c00703 |
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author | Werther, Philipp Yserentant, Klaus Braun, Felix Grußmayer, Kristin Navikas, Vytautas Yu, Miao Zhang, Zhibin Ziegler, Michael J. Mayer, Christoph Gralak, Antoni J. Busch, Marvin Chi, Weijie Rominger, Frank Radenovic, Aleksandra Liu, Xiaogang Lemke, Edward A. Buckup, Tiago Herten, Dirk-Peter Wombacher, Richard |
author_facet | Werther, Philipp Yserentant, Klaus Braun, Felix Grußmayer, Kristin Navikas, Vytautas Yu, Miao Zhang, Zhibin Ziegler, Michael J. Mayer, Christoph Gralak, Antoni J. Busch, Marvin Chi, Weijie Rominger, Frank Radenovic, Aleksandra Liu, Xiaogang Lemke, Edward A. Buckup, Tiago Herten, Dirk-Peter Wombacher, Richard |
author_sort | Werther, Philipp |
collection | PubMed |
description | [Image: see text] Small-molecule fluorophores enable the observation of biomolecules in their native context with fluorescence microscopy. Specific labeling via bio-orthogonal tetrazine chemistry combines minimal label size with rapid labeling kinetics. At the same time, fluorogenic tetrazine–dye conjugates exhibit efficient quenching of dyes prior to target binding. However, live-cell compatible long-wavelength fluorophores with strong fluorogenicity have been difficult to realize. Here, we report close proximity tetrazine–dye conjugates with minimal distance between tetrazine and the fluorophore. Two synthetic routes give access to a series of cell-permeable and -impermeable dyes including highly fluorogenic far-red emitting derivatives with electron exchange as the dominant excited-state quenching mechanism. We demonstrate their potential for live-cell imaging in combination with unnatural amino acids, wash-free multicolor and super-resolution STED, and SOFI imaging. These dyes pave the way for advanced fluorescence imaging of biomolecules with minimal label size. |
format | Online Article Text |
id | pubmed-8461766 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-84617662021-09-27 Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy Werther, Philipp Yserentant, Klaus Braun, Felix Grußmayer, Kristin Navikas, Vytautas Yu, Miao Zhang, Zhibin Ziegler, Michael J. Mayer, Christoph Gralak, Antoni J. Busch, Marvin Chi, Weijie Rominger, Frank Radenovic, Aleksandra Liu, Xiaogang Lemke, Edward A. Buckup, Tiago Herten, Dirk-Peter Wombacher, Richard ACS Cent Sci [Image: see text] Small-molecule fluorophores enable the observation of biomolecules in their native context with fluorescence microscopy. Specific labeling via bio-orthogonal tetrazine chemistry combines minimal label size with rapid labeling kinetics. At the same time, fluorogenic tetrazine–dye conjugates exhibit efficient quenching of dyes prior to target binding. However, live-cell compatible long-wavelength fluorophores with strong fluorogenicity have been difficult to realize. Here, we report close proximity tetrazine–dye conjugates with minimal distance between tetrazine and the fluorophore. Two synthetic routes give access to a series of cell-permeable and -impermeable dyes including highly fluorogenic far-red emitting derivatives with electron exchange as the dominant excited-state quenching mechanism. We demonstrate their potential for live-cell imaging in combination with unnatural amino acids, wash-free multicolor and super-resolution STED, and SOFI imaging. These dyes pave the way for advanced fluorescence imaging of biomolecules with minimal label size. American Chemical Society 2021-08-20 2021-09-22 /pmc/articles/PMC8461766/ /pubmed/34584958 http://dx.doi.org/10.1021/acscentsci.1c00703 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Werther, Philipp Yserentant, Klaus Braun, Felix Grußmayer, Kristin Navikas, Vytautas Yu, Miao Zhang, Zhibin Ziegler, Michael J. Mayer, Christoph Gralak, Antoni J. Busch, Marvin Chi, Weijie Rominger, Frank Radenovic, Aleksandra Liu, Xiaogang Lemke, Edward A. Buckup, Tiago Herten, Dirk-Peter Wombacher, Richard Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy |
title | Bio-orthogonal Red and Far-Red Fluorogenic Probes
for Wash-Free Live-Cell and Super-resolution Microscopy |
title_full | Bio-orthogonal Red and Far-Red Fluorogenic Probes
for Wash-Free Live-Cell and Super-resolution Microscopy |
title_fullStr | Bio-orthogonal Red and Far-Red Fluorogenic Probes
for Wash-Free Live-Cell and Super-resolution Microscopy |
title_full_unstemmed | Bio-orthogonal Red and Far-Red Fluorogenic Probes
for Wash-Free Live-Cell and Super-resolution Microscopy |
title_short | Bio-orthogonal Red and Far-Red Fluorogenic Probes
for Wash-Free Live-Cell and Super-resolution Microscopy |
title_sort | bio-orthogonal red and far-red fluorogenic probes
for wash-free live-cell and super-resolution microscopy |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8461766/ https://www.ncbi.nlm.nih.gov/pubmed/34584958 http://dx.doi.org/10.1021/acscentsci.1c00703 |
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