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Publications

Journal Article Epublish 5 citations

PubMed: 39463828 DOI: 10.1021/acscentsci.4c00616

Gentle Rhodamines for Live-Cell Fluorescence Microscopy.

ACS Cent Sci 10(10):1933-1944

Authors: Tianyan Liu, Julian Kompa, Jing Ling, Nicolas Lardon, Yuan Zhang, Jingting Chen, Luc Reymond, Peng Chen, Mai Tran, Zhongtian Yang, Haolin Zhang, Yitong Liu, Stefan Pitsch, Peng Zou, Lu Wang, Kai Johnsson, Zhixing Chen

Affiliations

  • College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China. Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, State Key Laboratory of Membrane Biology, Peking University, Beijing 100871, China.
  • Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg 69120, Germany.
  • College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China.
  • Biomolecular Screening Facility, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne 1015, Switzerland.
  • PKU-Nanjing Institute of Translational Medicine, Nanjing 211800, China. GenVivo Tech, Nanjing 211800, China.
  • Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, State Key Laboratory of Membrane Biology, Peking University, Beijing 100871, China.
  • Spirochrome AG, Chalberwiedstrasse 4, CH-8260 Stein am Rhein, Switzerland.
  • Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, State Key Laboratory of Membrane Biology, Peking University, Beijing 100871, China. College of Chemistry and Molecular Engineering, Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of the Ministry of Education, PKU-IDG/McGovern Institute for Brain Research, Peking University, Beijing 100871, China.
  • Key Laboratory of Smart Drug Delivery, Ministry of Education, School of Pharmacy, Fudan University, 201203 Shanghai, China.
  • College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China. Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, State Key Laboratory of Membrane Biology, Peking University, Beijing 100871, China. PKU-Nanjing Institute of Translational Medicine, Nanjing 211800, China. GenVivo Tech, Nanjing 211800, China.

Abstract

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.

MeSH

Cells, Microscopy, Fluorescence, Time-Lapse Imaging, Rhodamines, Microscopy

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