A microstructured antimicrobial film, an antimicrobial film assembly, a method of protecting a surface from microbial contamination, and a method of making a microstructured antimicrobial film. The microstructured antimicrobial film can include a substrate having a first side that includes a first major surface, a plurality of microstructured wells defined in the first side of the substrate, and an antimicrobial material positioned within at least some of the plurality of wells. Each of the plurality of wells can be at least partially defined by a base that is spaced a distance from the first major surface of the substrate. The antimicrobial material can be positioned, such that an upper surface of the antimicrobial material is spaced a distance from the first major surface of the substrate. The antimicrobial film assembly can include a roll or a stack of microstructured antimicrobial films.