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

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.

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3310733.pdf (Publisher version), 2MB
 
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 Creators:
Uno, K.1, Author              
Aktalay, A., Author
Bossi, M. L., Author
Irie, M., Author
Belov, V. N.2, Author              
Hell, S. W.1, Author              
Affiliations:
1Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society, ou_578627              
2Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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Free keywords: fluorescence; diarylethenes; bioconjugation; chromophores; Urbachtail
 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.

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Language(s): eng - English
 Dates: 2021-03-292021-04-06
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1073/pnas.2100165118
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Title: Proceedings of the National Academy of Sciences of the USA
Source Genre: Journal
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Pages: - Volume / Issue: 118 (14) Sequence Number: e2100165118 Start / End Page: - Identifier: -