Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy

Philipp Werther, Klaus Yserentant, Felix Braun, Kristin Grußmayer, Vytautas Navikas, Miao Yu, Zhibin Zhang, Michael J. Ziegler, Christoph Mayer, More Authors

Research output: Contribution to journalArticleScientificpeer-review

10 Citations (Scopus)
45 Downloads (Pure)


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.

Original languageEnglish
Pages (from-to)1561-1571
Number of pages11
JournalACS Central Science
Issue number9
Publication statusPublished - 2021


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