Gentle Rhodamines for Live-Cell Fluorescence Microscopy
Rhodamines have been continuously optimized in brightness, biocompatibility, and color 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.
Authors
- Tianyan Liu (ORCID: https://orcid.org/0000-0003-1982-2736)
- Zhixing Chen (ORCID: https://orcid.org/0000-0001-5962-7359)
- Peng Zou (ORCID: https://orcid.org/0000-0002-9798-5242)
- Stefan Pitsch (ORCID: https://orcid.org/0000-0002-9442-2727)
- Mai Tran (ORCID: https://orcid.org/0000-0003-1666-4584)
- Nicolas Lardon
- Kai Johnsson (ORCID: https://orcid.org/0000-0002-8002-1981)
- Jing Ling (ORCID: https://orcid.org/0000-0003-4620-9507)
- Zhongtian Yang (ORCID: https://orcid.org/0000-0002-9916-4616)
- Luc Reymond (ORCID: https://orcid.org/0000-0001-5282-9273)
- Peng R. Chen (ORCID: https://orcid.org/0000-0002-0402-7417)
- Julian Kompa (ORCID: https://orcid.org/0000-0002-4479-5429)
- Haolin Zhang (ORCID: https://orcid.org/0000-0002-9679-5850)
- Jingting Chen (ORCID: https://orcid.org/0000-0001-9806-9054)
- Lu Wang (ORCID: https://orcid.org/0000-0001-8412-2985)
- Yuan Zhang (ORCID: https://orcid.org/0000-0002-5534-5415)
- Yitong Liu (ORCID: https://orcid.org/0000-0003-4331-7422)
Institutions
- Peking University (CN)
- Fudan University (CN)
- Max Planck Institute for Medical Research (DE)
- Beijing National Laboratory for Molecular Sciences (CN)
- Center for Life Sciences (CN)
- École Polytechnique Fédérale de Lausanne (CH)
Publication Details
- Journal
- ACS Central Science
- Published
- 2024-10-02
- DOI
- https://doi.org/10.1021/acscentsci.4c00616
- Citations
- 20
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 13.25
Funders
- École Polytechnique Fédérale de Lausanne
- Fudan University
- Peking University
- Beijing Municipal Science and Technology Commission
- State Key Laboratory of Membrane Biology
- State Key Laboratory of Bio-organic and Natural Products Chemistry
- McGovern Institute for Brain Research, Massachusetts Institute of Technology
- School of Pharmacy, Fudan University
- Key Laboratory of Bioorganic Chemistry and Molecular Engineering
- Key Laboratory of Smart Drug Delivery
- National Key Research and Development Program of China
- Beijing National Laboratory for Molecular Sciences