Stainless steels are one of the most common materials used in health care environments. However, the lack of antibacterial advantage has limited their use in practical application. In this paper, antibacterial stainless steel surfaces with different Cu contents have been prepared by plasma surface alloying technology (PSAT). The steel surface with Cu content 90 wt.% (Cu-SS) exhibits strong bactericidal activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) within 3 h. Although the Cu-containing surface with Cu content 2.5 wt.% (CuNi-SS) can also kill all tested bacteria, this process needs 12 h. SEM observation of the bacterial morphology and an agarose gel electrophoresis were performed to study the antibacterial mechanism of Cu-containing stainless steel surfaces against E. coli. The results indicated that Cu ions are released when the Cu-containing surfaces are in contact with bacterial and disrupt the cell membranes, killing the bacteria. The toxicity of Cu-alloyed surfaces does not cause damage to the bacterial DNA. These results provide a scientific explanation for the antimicrobial applications of Cu-containing stainless steel. The surfaces with different antibacterial abilities could be used as hygienic surfaces in healthcare-associated settings according to the diverse requirement of bactericidal activities. (C) 2012 Elsevier B.V. All rights reserved.