Fluorescent microspheres were easily fabricated from aqueous CdTe nanocrystals (NCs). The NCs, which had negative charges on the surface, were first extracted to chloroform by cationic surfactant octadecyl-p-vinyl-benzyldimethylammonium chloride (OVDAC) and then swollen into performed polystyrene (PS) microspheres. Through this method, strong photoluminescence (PL) of aqueous NCs was inherited in the resultant composite microspheres. Moreover, the NCs were firmly stabilized in the microspheres, withstanding not only polar solvents but also nonpolar solvents. UV-vis spectrum, PL spectrum, TEM, and confocal fluorescence microscopy were used to characterize the product.