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Calcein was first covalently grafted onto the surface of magnetic silica nanoparticles to obtain a novel fluorescent nanoprobe for sensitive and selective determination of Cu(II) ion. The nanoparticles were synthesized in a two-step reaction under mild conditions and characterized using small-angle Xray scattering, Fourier transform infrared spectroscopy, transmission electron microscopy, and thermogravimetric analysis. The nanoprobe also displays good selectivity over other metal ions and is highly stable under conditions of high ionic strength. The response to Cu(II) ranges from 80 nM to 1.25 μM. The detection limit (LOD) is 43 nM which is lower than previously reported LODs. The probe can be regenerated by adding the complexing agent EDTA and also can be separated from a sample mixture by applying a magnetic field. We conclude that this method represents a new and convenient approach to the development of magnetic and reusable optical probes.