Fabrication and characterization of YVO4:Eu3+ nanophosphors prepared by microwave (MW) irradiation assisted soft template synthesis were reported. The effects of synthesis conditions such as different powers of MW irradiation, pH values and concentration of reaction materials on properties of nanophosphor were also investigated to obtain the controllable size, morphology and high luminescence efficiency. Morphology, crystalline structure, and optical properties were characterized by field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD) and fluorescence spectroscopy, respectively. The results showed that YVO4:Eu3+ nanophosphors were obtained by using diethyleneglycol (DEG) as soft template, with pH values in the range of 4 to 12, upon microwave irradiation from 300 to 900 W, at temperature of 80 °C. The high fluorescent YVO4:Eu3+ nanocrystals obtained with size from 15 nm down to 8 nm are more effective to develop an ultrahigh sensitive fluorescent label for biomolecule, cell and tissue.