Polyacrylamide microspheres have been successfully used to reduce water production in reservoirs, but it is impossible to distinguish polyacrylamide microspheres from polyacrylamide that is used to enhance oil recovery and is already present in production fluids. In order to detect polyacrylamide microspheres in the reservoir produced fluid, fluorescent polyacrylamide microspheres P(AM-BA-AMCO), which fluoresce under ultraviolet irradiation, were synthesized via an inverse suspension polymerization. In order to keep the particle size distribution in a narrow range, the synthesis conditions of the polymerization were studied, including the stirring speed and the concentrations of initiator, Na2CO3, and dispersant. The bonding characteristics of microspheres were determined by Fourier transform infrared spectroscopy. The surface morphology of these microspheres was observed under ultraviolet irradiation with an inverse fluorescence microscope. A laboratory evaluation test showed that the fluorescent polymer microspheres had good water swelling capability, thus they had the ability to plug and migrate in a sand pack. The plugging rate was 99.8 % and the residual resistance coefficient was 800 after microsphere treatment in the sand pack. Furthermore, the fluorescent microspheres and their fragments were accurately detected under ultraviolet irradiation in the produced fluid, even though they had experienced extrusion and deformation in the sand pack.