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A novel, successful method of bactericidal treatment of pathogenic bacterial biofilms in vitro by laser-induced forward transfer of metallic nanoparticles from a polyethylene terephthalate polymeric substrate was suggested. Transferred nanoparticles were characterized by scanning and transmission electron microscopy, energy-dispersive X-ray and Raman spectroscopy. The antibacterial modality of the method was tested on Gram-positive (<i>Staphylococcus aureus</i>) and Gram-negative (<i>Pseudomonas Aeruginosa</i>) bacterial biofilms in vitro, revealing their complete destruction. The proposed simple, cost-effective and potentially mobile biofilm treatment method demonstrated its high and broad bactericidal efficiency.