A simple and convenient strategy has been put forward to fabricate smart fluorescent magnetic wheat germ agglutinin-modified trifunctional nanospheres (WGA-TFNS) for recognition of human prostate carcinoma DU-145 cells which are surface-expressed with sialic acid and N-acetylglucosamine. These TFNS can be easily manipulated, tracked, and conveniently used to capture and separate target cells. The presence of wheat germ agglutinin on the surface of WGA-TFNS was confirmed by FTIR, biorecognition of carboxymethyl chitin-modified quantum dots (CM-CT-QDs), and bacterium Staphylococcus aureus. The success in recognizing DU-145 cells by the WGA-TFNS indicates that WGA-TFNS could be applicable.