A series of novel hybrid compounds was synthesized where 2 '-hydroxyacetophenone and N-alkylated thiotetrazole were coupled via methylene spacers of different lengths. The structures of the compounds were characterized by spectroscopic techniques, including the 1H-and 13C-NMR, FT-IR, and HRMS methods. The antimicrobial activities of the title compounds were evaluated against clinical isolates of Gram-positive and negative bacterial and fungal species. All compounds showed a broad spectrum of antimicrobial activity and inhibited all bacteria and fungi that tested with MIC values ranging from 4 to 128 mu g/ml. 4a and 5d were the most active antibacterial members.