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FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors
Published on: August 20, 2012
A reversible fluorescence chemosensor for sequentially quantitative monitoring copper and sulfide in living cells
Qingtao Meng1, Run Zhang2, Hongmin Jia1
1School of Chemical Engineering, University of Science and Technology Liaoning, Anshan 114044, China.
Abstract:
We report a novel, selective and sensitive strategy for the sequentially "ON-OFF-ON" fluorescent detection of Cu(2+) and S(2-) based on a fluorescein derivative, FL. The specific binding of FL towards Cu(2+) in aqueous and biological media led to the intensive green fluorescence quenching and a notable increase of the absorbance maximum at 480 nm. In the presence of S(2-), the intensity and overall pattern of the fluorescence emission and UV-vis spectra of FL-Cu(2+) ensemble were recovered since the abolishment of paramagnetic Cu(2+). This displacement approach exhibited highly specificity, and sensitivity with detection limits of 3 nM for Cu(2+) and 150 nM for S(2-). The fluorescence "ON-OFF-ON" circle can be repeated to a minimum of 5 times by the alternative addition of Cu(2+) and S(2-), implying that FL is a renewable dual-functional chemosensor. The biocompatibility of FL toward breast carcinoma cells, MDA-MB-231 was confirmed by MTT assay. The reversible "ON-OFF-ON" fluorescent response of FL to Cu(2+) and S(2-) in living system was further confirmed by confocal fluorescence imaging of living cells. The quantification of Cu(2+) and S(2-) in single intact cell was realized by the flow cytometry analysis.

