Based on the self-ordered ring(SOR) fluorescence microscopic imaging technique on a hydrophobic glass slide with Zn 2+ and cetyltrimethyl ammonium bromide(CTMAB) as sensitizer,and poly(vinyl alcohol)-124(PVA-124) and NH 3-NH 4 Cl(pH 10.00) as the medium,a method has been developed for determining sparfloxacin(SPFX) concentrations in chicken serum and residues in chicken tissues and manures.When the droplet volume was 0.20 μ L,SPFX was determined in the range of 1.38×10-13 ~2.03×10-12 mol.ring-1(or 6.90×10-7 ~1.02×10-5 mol.L-1),and the limit of detection(LOD) was 14fmol.ring-1(or 6.90× 10-8 mol.L-1).The recoveries of SPFX at all different spiked levels are in the range of 90.74%~106.61% when the methanol or acetonitrile were used as extracting agent,respectively,and the relative standard deviations(RSDs) are less than 3.0%.This study expands the applied fields of SOR technique in drug concentrations and residues determination.