Print Email Facebook Twitter 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 Author Werther, Philipp (University of Heidelberg) Yserentant, Klaus (University of Heidelberg) Braun, Felix (University of Heidelberg) Grußmayer, K.S. (TU Delft BN/Kristin Grussmayer Lab; TU Delft BN/Afdelingsbureau; Swiss Federal Institute of Technology; Kavli institute of nanoscience Delft) Navikas, Vytautas (Swiss Federal Institute of Technology) Yu, Miao (Johannes Gutenberg-University Mainz) Zhang, Zhibin (University of Heidelberg; Harbin Institute of Technology) Ziegler, Michael J. (University of Heidelberg; Max Planck Institute for Medical Research) Mayer, Christoph (University of Heidelberg) Date 2021 Abstract 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. To reference this document use: http://resolver.tudelft.nl/uuid:d96d5236-c2b8-4b51-8b9d-b367bb35536b DOI https://doi.org/10.1021/acscentsci.1c00703 ISSN 2374-7943 Source ACS Central Science, 7 (9), 1561-1571 Part of collection Institutional Repository Document type journal article Rights © 2021 Philipp Werther, Klaus Yserentant, Felix Braun, K.S. Grußmayer, Vytautas Navikas, Miao Yu, Zhibin Zhang, Michael J. Ziegler, Christoph Mayer, More Authors Files PDF acscentsci.1c00703.pdf 6.67 MB Close viewer /islandora/object/uuid:d96d5236-c2b8-4b51-8b9d-b367bb35536b/datastream/OBJ/view