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  Gentle rhodamines for live-cell fluorescence microscopy

Liu, T., Kompa, J., Ling, J., Lardon, N., Zhang, Y., Chen, J., et al. (2024). Gentle rhodamines for live-cell fluorescence microscopy. bioRxiv. doi:10.1101/2024.02.06.579089.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000F-DE00-7 Version Permalink: https://hdl.handle.net/21.11116/0000-000F-DE01-6
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 Creators:
Liu, Tianyan, Author
Kompa, Julian1, Author           
Ling, Jing, Author
Lardon, Nicolas1, Author           
Zhang, Yuan, Author
Chen, Jingting, Author
Reymond, Luc, Author
Chen, Peng, Author
Tran, Mai1, Author           
Yang, Zhongtian, Author
Zhang, Haolin, Author
Liu, Yitong, Author
Pitsch, Stefan, Author
Zou, View ORCID ProfilePeng, Author
Wang, Lu, Author
Johnsson, Kai1, Author           
Chen, Zhixing, Author
Affiliations:
1Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society, ou_2364732              

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 Abstract: Rhodamines have been continuously optimized in brightness, biocompatibility, and colors to fulfill the demands of modern bioimaging. However, the problem of phototoxicity caused by the excited fluorophore under long-term illumination has been largely neglected, hampering their use in time-lapse imaging. Here we introduce cyclooctatetraene (COT) conjugated rhodamines that span the visible spectrum and exhibit significantly reduced phototoxicity. We identified a general strategy for the generation of Gentle Rhodamines, which preserved their outstanding spectroscopic properties and cell permeability while showing an efficient reduction of singlet-oxygen formation and diminished cellular photodamage. Paradoxically, their photobleaching kinetics do not go hand in hand with reduced phototoxicity. By combining COT-conjugated spirocyclization motifs with targeting moieties, these gentle rhodamines compose a toolkit for time-lapse imaging of mitochondria, DNA, and actin and synergize with covalent and exchangeable HaloTag labeling of cellular proteins with less photodamage than their commonly used precursors. Taken together, the Gentle Rhodamines generally offer alleviated phototoxicity and allow advanced video recording applications, including voltage imaging.

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Language(s): eng - English
 Dates: 2024-02-08
 Publication Status: Published online
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 Identifiers: DOI: 10.1101/2024.02.06.579089
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Title: bioRxiv
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