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Related Experiment Video

Updated: Jun 17, 2026

Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein
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Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein

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Visualizing ascorbate-triggered release of labile copper within living cells using a ratiometric fluorescent sensor.

Dylan W Domaille1, Li Zeng, Christopher J Chang

  • 1Department of Chemistry and the Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA.

Journal of the American Chemical Society
|January 8, 2010
PubMed
Summary
This summary is machine-generated.

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Researchers developed Ratio-Coppersensor-1 (RCS1), a novel fluorescent sensor for imaging copper ions in living cells. This sensor offers high selectivity for Cu(+) and enables ratiometric imaging of labile copper pools in kidney and brain cells.

Area of Science:

  • Chemical Biology
  • Biomedical Engineering
  • Molecular Imaging

Background:

  • Copper ions (Cu(+)) play crucial roles in cellular processes, but their dysregulation is linked to various diseases.
  • Accurate imaging of labile copper pools in living cells is challenging due to the need for selective and sensitive probes.

Purpose of the Study:

  • To synthesize and characterize Ratio-Coppersensor-1 (RCS1), a novel water-soluble fluorescent sensor for ratiometric imaging of copper ions in living cells.
  • To evaluate the selectivity, sensitivity, and applicability of RCS1 for sensing labile Cu(+) in cellular environments.

Main Methods:

  • Synthesis of RCS1, a sensor combining a BODIPY reporter and a thioether ligand.
  • Characterization of spectroscopic properties and metal ion selectivity (Cu(+) vs. Cu(2+), Zn(2+)).

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  • Live-cell confocal microscopy to assess membrane permeability, ratiometric imaging capabilities, and cellular copper dynamics.

    • Main Results:

    • RCS1 exhibits high selectivity and sensitivity for Cu(+) over other biologically relevant metal ions.
    • Cu(+) binding induces a significant fluorescence ratio change (approx. 20-fold) with visible excitation/emission profiles.
    • RCS1 successfully performed ratiometric imaging of labile Cu(+) pools in kidney and brain cells, detecting changes triggered by ascorbate stimulation.

    Conclusions:

    • RCS1 is a valuable tool for ratiometric imaging of labile copper in living cells.
    • The sensor's properties facilitate the study of copper homeostasis and its role in cellular function and disease.
    • RCS1 enables real-time monitoring of intracellular copper dynamics in response to physiological stimuli.