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  A general design of caging-group-free photoactivatable fluorophores for live-cell nanoscopy

Lincoln, R., Bossi, M. L., Remmel, M., D'Este, E., Butkevich, A. N., & Hell, S. W. (2022). A general design of caging-group-free photoactivatable fluorophores for live-cell nanoscopy. Nature Chemistry, 14(9), 1013-1020. doi:10.1038/s41557-022-00995-0.

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アイテムのパーマリンク: https://hdl.handle.net/21.11116/0000-000B-3E96-7 版のパーマリンク: https://hdl.handle.net/21.11116/0000-000C-3B30-C
資料種別: 学術論文

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s41557-022-00995-0.pdf (出版社版), 4MB
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3400985.pdf (プレプリント), 6MB
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作成者

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 作成者:
Lincoln, Richard, 著者
Bossi, Mariano Luis1, 著者           
Remmel, Michael, 著者
D'Este, Elisa, 著者
Butkevich, Alexey N., 著者
Hell, Stefan W.1, 著者                 
所属:
1Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, ou_3350048              

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 要旨: The controlled switching of fluorophores between non-fluorescent and fluorescent states is central to every super-resolution fluorescence microscopy (nanoscopy) technique, and the exploration of radically new switching mechanisms remains critical to boosting the performance of established, as well as emerging super-resolution methods. Photoactivatable dyes offer substantial improvements to many of these techniques, but often rely on photolabile protecting groups that limit their applications. Here we describe a general method to transform 3,6-diaminoxanthones into caging-group-free photoactivatable fluorophores. These photoactivatable xanthones (PaX) assemble rapidly and cleanly into highly fluorescent, photo- and chemically stable pyronine dyes upon irradiation with light. The strategy is extendable to carbon- and silicon-bridged xanthone analogues, yielding a family of photoactivatable labels spanning much of the visible spectrum. Our results demonstrate the versatility and utility of PaX dyes in fixed and live-cell labelling for conventional microscopy, as well as the coordinate-stochastic and deterministic nanoscopies STED, PALM and MINFLUX.

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言語: eng - English
 日付: 2022-07-212022-09
 出版の状態: 出版
 ページ: -
 出版情報: -
 目次: -
 査読: 査読あり
 識別子(DOI, ISBNなど): DOI: 10.1038/s41557-022-00995-0
 学位: -

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Project name : The research was funded by Bundesministerium für Bildung und Forschung (German Federal Ministry of Education and Research), project no. 13N14122 ‘3D Nano Life Cell’ (S.W.H.). R.L. is grateful to the Max Planck Society for a Nobel Laureate Fellowship. We thank S. Jakobs (MPI-NAT) for providing the U2OS-Vim-Halo and U2OS-Vim-SNAP cells and the EMBL for providing the U2OS-ZFN-SNAP-Nup107, U2OS-NUP96-Halo and HK-2xZFN-mEGFP-Nup107 cells. We thank the following people at the MPI for Medical Research: S. Fabritz (Mass Spectrometry Core Facility) for recording mass spectra of proteins and small molecules, M. Steigleder (Electronics Workshop) for building the custom 405-nm light source for the Penn m1 photoreactor, K. Johnsson (Department of Chemical Biology) for the SNAP-tag protein, M. Tarnawski (Protein Expression and Characterization Facility) for the HaloTag7 protein, and our colleagues at the Department of Optical Nanoscopy—J. Hubrich for supporting the cell culture, A. Aktalay for assistance with labelling of nanobody samples, and J. Matthias for critical reading of the manuscript and advice on sample preparation for MINFLUX. We thank the team at Abberior Instruments for support on the MINFLUX image acquisition.
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出版物 1

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出版物名: Nature Chemistry
  省略形 : Nat. Chem.
種別: 学術雑誌
 著者・編者:
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出版社, 出版地: London, UK : Nature Publishing Group
ページ: - 巻号: 14 (9) 通巻号: - 開始・終了ページ: 1013 - 1020 識別子(ISBN, ISSN, DOIなど): ISSN: 1755-4330
CoNE: https://pure.mpg.de/cone/journals/resource/1755-4330