Metallic glass as a prominent class of structure and multifunctional materials exhibits several unique properties in mechanical, electrochemical, and thermal properties. This study aimed to realize the advantage of biomedical application and to promote the attainable size of metallic glasses by the physical vapor deposition. The Zr-Cu-Al-Ag thin film metallic glass (TFMG) was deposited on silicon wafer and SUS304 stainless steel substrates by magnetron sputtering with single target. For X-ray diffraction analysis, all TFMGs revealed typical broad peaks around the incident angle of 30 to 50, suggesting that coatings possess amorphous structure. In addition, diffuse halo ring patterns of transmission electron microscopy indicated a fine amorphorization for TFMG via sputtering process. The variation of surface roughness showed that TFMG derived from higher power of metallic targets revealed rougher morphology. Besides, the roughness of SUS304 stainless steel substrate significantly reduced from 7 nm to about 1 nm after TFMGs were deposited. The microbes of Candida albicans, Escherichia coli, and Pseudomonas aeruginosa were used and cultivated on the TFMG coatings with medium to investigate the antimicrobial properties. In the incubation experiment the growth of each microbe was recorded by a digital photography system and the growth area was calculated by image processing software. The growth area of the microbes on the TFMG was mostly smaller than that on SUS304 stainless steel ones within incubation time of 72 h, indicating that the TFMGs reveal better antimicrobial capability. Moreover, the coatings exhibit a particularly long-term antimicrobial effect for P. aeruginosa. In summary, the Zr-Cu-Al-Ag prepared by sputtering with a single target device presented superior glass forming ability, and coatings with copper and silver constituents revealed significantly antimicrobial properties. Besides, the surface roughness is another factor to affect the biological activities, suggesting that microbes are difficult to attach and move on an ultra-smooth surface. It is demonstrated in this study that the TFMGs extend attainable size of the metallic glass and show a potential application in medical related field. (C) 2013 Elsevier B.V. All rights reserved.