A self-ordered ring (SOR) fluorescence microscopic imaging technique has been developed for the determination of trace amounts of sulfadiazine based on its derivatization with fluorescamine. In the presence of HAc-NaAc buffer solution (pH 3.12) and polyvinyl alcohol-124 (PVA-124), the droplet containing fluorescamine derivatized sulfadiazine can form a SOR on the solid support after solvent evaporation with the diameter of 1.86 mm and its ring belt width of 54.9 μm. The quantitative analysis of sulfadiazine is achieved with the linear range of 7.8×10-14～1.8×10-12 mol·ring-1 (3.9×10-7～9.0×10-6 mol·L-1) and detection limit of 7.8×10-15 mol·ring-1 (3.9×10-8 mol·L-1) when 0.2 μL droplet was spotted. The technique has been satisfactorily applied to the determination of sulfadiazine in the tablet, synthetic sample and residues in six different milk samples with the recoveries of 91.0%～105.8%, respectively, and RSDs less than 4.4%.