The use of radiopharmaceuticals is a good way to track and treat infections. In this study, Gemifloxacin a broad-spectrum antibiotic from the family of fluoroquinolones by using of Technetium^99m (Tc^99m) and direct method was labeled and used. Gemifloxacin by using of technetium^99m and direct method was labeled, then the stability of the drug, bind to Escherichia coli and bio-distribution in Mice was examined then possible changes in tissue by Hematoxylin and eosin (H&E) staining and immunohistochemical (IHC) staining for Desmin marker was evaluated. By using of High performance liquid chromatography (HPLC), labeling efficiency, 96.94 % and binding to bacteria 95.43 % was determined. The results of radiopharmaceutical biodistributions indicate its tendency to the position of infection. Presence of lesions in the muscle tissues after infection was confirmed by histopathological methods and since excretion of drug, effects diminished. Desmin was shown in all samples. The combination can be used as a factor in taking pictures of places with Escherichia coli infections, without causing tissue damage.