In the present investigation an attempt has been made to develop efficient, rapid, three-component, one-pot synthesis of 2,3-disubstituted thiazolidin-4-one derivatives under different reaction conditions like reflux, stirring and microwave irradiation approach. All the synthesized compounds were characterized by melting point, elemental analysis, IR, 1H NMR and Mass spectroscopy. Further, all the synthesized compounds were screened in vitro for their antimicrobial activities in terms of zone of inhibition and minimum inhibitory concentration.