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Journal Article

Turn-on mode diarylethenes for bioconjugation and fluorescence microscopy of cellular structures

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Uno,  K.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

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Belov,  V. N.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Hell,  S. W.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

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Citation

Uno, K., Aktalay, A., Bossi, M. L., Irie, M., Belov, V. N., & Hell, S. W. (2021). Turn-on mode diarylethenes for bioconjugation and fluorescence microscopy of cellular structures. Proceedings of the National Academy of Sciences of the USA, 118(14): e2100165118. doi:10.1073/pnas.2100165118.


Cite as: http://hdl.handle.net/21.11116/0000-0008-4CAD-1
Abstract
The use of photoswitchable fluorescent diarylethenes (fDAEs) as protein labels in fluorescence microscopy and nanoscopy has been limited by labeling inhomogeneity and the need for ultraviolet light for fluorescence activation (on-switching). To overcome these drawbacks, we prepared “turn-on mode” fDAEs featuring thienyl substituents, multiple polar residues, and a reactive maleimide group in the core structure. Conjugates with antibodies and nanobodies displayed complete on-switching and excitation with violet (405 nm) and yellow-green (<565 nm) light, respectively. Besides, they afforded high signal-to-noise ratios and low unspecific labeling in fluorescence imaging. Irradiation with visible light at 532 nm or 561 nm led to transient on-off switching (“blinking”) of the fDAEs of double-labeled nanobodies so that nanoscale superresolution images were readily attained through switching and localization of individual fluorophores.