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Photoactivatable fluorophore for stimulated emission depletion (STED) microscopy and bioconjugation technique for hydrophobic labels

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Bossi,  Mariano
Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society;

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Hell,  Stefan W.
Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society;

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Citation

Weber, M., Khan, T. A., Patalag, L. J., Bossi, M., Leutenegger, M., Belov, V. N., et al. (2021). Photoactivatable fluorophore for stimulated emission depletion (STED) microscopy and bioconjugation technique for hydrophobic labels. Chemistry – A European Journal, 27(1), 451-458. doi:10.1002/chem.202004645.


Cite as: http://hdl.handle.net/21.11116/0000-0007-4D36-7
Abstract
The use of photoactivatable dyes in STED microscopy has so far been limited by two—photon activation through the STED beam and by the fact that photoactivatable dyes are poorly solvable in water. Here we report ONB‐2SiR, a fluorophore that can be both photoactivated in the UV and specifically de‐excited by STED at 775 nm. Likewise, we introduce a conjugation and purification protocol to effectively label primary and secondary antibodies with moderately water‐soluble dyes. Greatly reducing dye aggregation, our technique provides a defined and tunable degree of labeling, and improves the imaging performance of dye conjugates in general.