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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 FRET-based fluorescent probe for mercury ions in water and living cells
Bo Zhang1, Pinyi Ma1, Dejiang Gao2
1College of Chemistry, Jilin University, Qianjin Street 2699, Changchun 130012, China.
Abstract:
On the basis of fluorescence resonance energy transfer (FRET), a new rhodamine derivative (DRh) was synthesized as a ratiometric fluorescent probe for detecting Hg(2+) in water and living cells samples. The recognition properties of the probe DRh with metal ions had been investigated in H2O/CH3CN (9:1, v/v; Tris-HCl 50mmolL(-1); pH=7.0) solution by the UV-Vis spectrophotometry and the fluorescence spectrophotometry. The results showed that the probe DRh exhibited the selective recognition of Hg(2+). Upon the addition of Hg(2+), the spirolactam ring of probe DRh was opened. The 1:1 stoichiometric structure between DRh and Hg(2+) were supported by Job's plot, MS and DFT theoretical calculations. The linearly fluorescence intensity ratio (I582/I538) is proportional to the concentration of Hg(2+) in the range 0-30μmolL(-1). The limit of detection (LOD) of Hg(2+) is 0.008μmolL(-1) (base on S/N=3). The present probe was applied to the determination of Hg(2+) in neutral water samples and gave recoveries ranging from 104.5 to 107.9%. Furthermore, the fluorescent probe also can be applied as a bioimaging reagent for Hg(2+) detection in HeLa cells.
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