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Journal Article

Bioorthogonal Caging-Group-Free Photoactivatable Probes for Minimal-Linkage-Error Nanoscopy

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Bossi,  Mariano Luis
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Hell,  Stefan W.       
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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acscentsci.3c00746.pdf
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Citation

Aktalay, A., Lincoln, R., Heynck, L., Lima, M. A. d. R. B. F., Butkevich, A. N., Bossi, M. L., et al. (2023). Bioorthogonal Caging-Group-Free Photoactivatable Probes for Minimal-Linkage-Error Nanoscopy. ACS Central Science, 9(8), 1581-1590. doi:10.1021/acscentsci.3c00746.


Cite as: https://hdl.handle.net/21.11116/0000-000D-BE6A-7
Abstract
Here we describe highly compact, click compatible, and photoactivatable dyes for super-resolution fluorescence microscopy (nanoscopy). By combining the photoactivatable xanthone (PaX) core with a tetrazine group, we achieve minimally sized and highly sensitive molecular dyads for the selective labeling of unnatural amino acids introduced by genetic code expansion. We exploit the excited state quenching properties of the tetrazine group to attenuate the photoactivation rates of the PaX, and further reduce the overall fluorescence emission of the photogenerated fluorophore, providing two mechanisms of selectivity to reduce the off-target signal. Coupled with MINFLUX nanoscopy, we employ our dyads in the minimal-linkage-error imaging of vimentin filaments, demonstrating molecular-scale precision in fluorophore positioning.