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The fluorescent polymer microsphere is a newly developed chemical agent for conformance control in reservoirs. In this paper, one kind of fluorescent polymer microspheres P(AM-BA-RhB) was synthesized via the inverse suspension polymerization method with Rhodamine B as a fluorescence functional monomer. Laboratory experiments were performed to characterize the morphology, fluorescent property, swelling property and plugging behavior of fluorescent polymer microspheres. The experimental results showed that the polymer microspheres P(AM-BA-RhB) displayed stable fluorescence performance in solutions containing metal ions at pH values between 3.0 and 10.0. The swelling property was not dramatically affected by the Rhodamine B embedded in the polymer microspheres by grafting. Both a visual micromodel test and sand-pack tubes experiment demonstrated that the fluorescent polymer microspheres could pass directly or by deformation through porous media and get into the in-depth formation. The injection pressure showed the phenomenon of "Wave-type Variation". Three plugging behaviors such as piston plugging, protruding plugging and fingering plugging were put forward. The introduction of fluorescent polymer microspheres could provide one method to research the conformance control and EOR mechanism of polymer microspheres in the reservoirs.