An antimicrobial substrate (glass, ceramic or metallic) coated on at least one of its surface with at least one mixed layer deposited by a sputtering under vacuum magnetically enhanced process is described. The layer comprising at least one antimicrobial agent mixed to binder material chosen amongst the metal oxides, oxynitrides, oxycarbides or nitrides. This substrate present antimicrobial properties, in particular bactericidal activity even when no thermal treatment has been applied. If a tempered and antimicrobial glass is required, the same co-sputtering process can be used, optionally an underlayer can be added. Antimicrobial properties are maintained even after a tempering process.