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

ABSTRACT: We present the synthesis, properties, and biological applications of Ratio-Coppersensor-1 (RCS1), a new water-soluble fluorescent sensor for ratiometric imaging of copper in living cells. RCS1 combines an asymmetric BODIPY reporter and thioether-based ligand receptor to provide high selectivity and sensitivity for Cu(+) over other biologically relevant metal ions, including Cu(2+) and Zn(2+), a ca. 20-fold fluorescence ratio change upon Cu(+) binding, and visible excitation and emission profiles compatible with standard fluorescence microscopy filter sets. Live-cell confocal microscopy experiments show that RCS1 is membrane-permeable and can sense changes in the levels of labile Cu(+) pools within living cells by ratiometric imaging, including expansion of endogenous stores of exchangeable intracellular Cu(+) triggered by ascorbate stimulation in kidney and brain cells.

SUBMITTER: Domaille DW

PROVIDER: S-EPMC2819030 | biostudies-literature | 2010 Feb

REPOSITORIES: biostudies-literature

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

Domaille Dylan W DW Domaille Dylan W DW Zeng Li L Chang Christopher J CJ

Journal of the American Chemical Society 20100201 4

We present the synthesis, properties, and biological applications of Ratio-Coppersensor-1 (RCS1), a new water-soluble fluorescent sensor for ratiometric imaging of copper in living cells. RCS1 combines an asymmetric BODIPY reporter and thioether-based ligand receptor to provide high selectivity and sensitivity for Cu(+) over other biologically relevant metal ions, including Cu(2+) and Zn(2+), a ca. 20-fold fluorescence ratio change upon Cu(+) binding, and visible excitation and emission profiles ...[more]

PMID: 20052977

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