Live‐cell localization microscopy with a fluorogenic and self‐blinking tetrazine probe

Philipp Werther, Klaus Yserentant, Felix Braun , Nicolai Kaltwasser, Christoph Popp , Mathis Baalmann , Dirk-Peter Herten, Richard Wombacher

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)
218 Downloads (Pure)


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.
Original languageEnglish
Pages (from-to)804-810
Number of pages7
JournalAngewandte Chemie - International Edition
Issue number2
Early online date22 Oct 2019
Publication statusPublished - 7 Jan 2020


  • bioorthogonal chemistry
  • click chemistry
  • fluorescent probes
  • super-resolution imaging
  • tetrazines

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)


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