Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 59, Issue 2, Pages 804-810Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201906806
Keywords
bioorthogonal chemistry; click chemistry; fluorescent probes; super-resolution imaging; tetrazines
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Funding
- Deutsche Forschungsgemeinschaft DFG [SPP1623, WO 1888/1-2]
- Federal Ministry of Education and Research (BMBF/VDI)
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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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