Thiazolidine-2,4-dione derivatives (4a-4e) have been synthesized and characterized by physico-chemical, elemental (C,H,N,S), FT-IR, mass and 1HNMR spectral analysis. In this context, the aim was to synthesized thiazolidine-2,4-dione derivatives and were screened for antimicrobial activity. Antimicrobial activity was screened using agar well diffusion assay method for determination of the diameter of zone of inhibition in millimeter (mm) and solid dilution method for determination of minimum inhibitory concentration (MIC) in µg/ml against Gram-positive(Staphylococcus aureus MTCC 1430, Bacillus subtilis MTCC 0441), Gram- negative(Escherichia coli MTCC 1573, Pseudomonas aeruginosa MTCC 2453) and fungal strains (Aspergillus tubingensis MTCC 2546). The antimicrobial activity of compound 4c2-(4-((2,4-dioxothiazolidin-5-ylidene)methyl)phenoxy)-N-(4-fluorophenyl)acetamidewas found to be most potent against all the tested strains of microorganisms with the zone of inhibition 17.1-19.5 mm at 200µg/ml and minimum inhibitory concentration value 0.6-0.8 µg/ml.