Green Fluorescent Protein GFP-Chromophore-Based Probe for the Detection of Mitochondrial Viscosity in Living Cells

Lei Cai, Huan Li, Xiang Yu, Luling Wu, Xiaoqin Wei, Tony D. James, Chusen Huang

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Viscosity is a pivotal factor for indicating the dysfunction of the mitochondria. To date, most of the fluorescent probes developed for mitochondrial viscosity have been designed using BODIPY, hemicyanine, or pyridine-based molecular rotors as part of the core structure. Our aim with this research was to extend the range of suitable fluorophores available for the construction of such fluorescent molecular rotors for evaluating the viscosity of mitocondria. Herein, we have developed a green fluorescent protein (GFP)-chromophore-based fluorescent probe (MIT-V) for the detection of mitochondrial viscosity in live cells. MIT-V exhibited a high sensitivity toward viscosity (from 7.9 cP to 438.4 cP). The "off-on"sensing mechanism of MIT-V was ascribed to the restricted rotation of single bonds and excited-state C= C double bonds of MIT-V. Cell studies indicated that MIT-V targets the mitochondria and that it was able to monitor real-time changes in the viscosity of live HeLa cell mitochondria. Therefore, we propose that MIT-V can be used as an effective chemosensor for the real-time imaging of mitochondrial viscosity in live cells. Our results clearly demonstrate the utility of such GFP-chromophore-based derivatives for the development of viscosity-sensitive systems.

Original languageEnglish
Pages (from-to)2128-2134
Number of pages7
JournalACS Applied Bio Materials
Issue number3
Early online date14 Jan 2021
Publication statusPublished - 15 Mar 2021

Bibliographical note

Funding Information:
C.H. thanks the National Natural Science Foundation of China (Grants 21672150, 21302125), Alexander von Humboldt Foundation (AvH), Shanghai Rising-Star Program (19QA1406400), Shanghai Government (18DZ2254200), Program for Changjiang Scholars and Innovative (IRT_16R49). T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University for support (2020ZD01). L.W. wishes to thank the China Scholarship Council and the University of Bath for supporting his PhD in the United Kingdom.


  • fluorescent probe
  • GFP-chromophore
  • live cell imaging
  • mitochondria
  • viscosity

ASJC Scopus subject areas

  • Biomaterials
  • Chemistry(all)
  • Biomedical Engineering
  • Biochemistry, medical


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