Cotton-fabric-supported silver nanoparticles (Ag NPs) have aroused great attention due to their remarkable physical and chemical properties and excellent broad-spectrum antibacterial performance.In this work, a surface dielectric barrier discharge (DBD) plasma method is developed and employed to prepare cotton fabric supported Ag NPs (Ag/cotton) for the first time. UV-Vis and X-ray photoelectron spectroscopy (XPS) results confirm the formation of Ag NPs. TEM images show that the size of Ag NPs is in the range 4.8−5.3 nm. Heat-sensitive cotton fabrics are not destroyed by surface DBD plasma according to FTIR and XRDresults. Wash fastness of the Ag/cotton samples is investigated using ultrasonic treatment for 30 min and it is shown that the Ag NPs possess good adhesion to the cotton fabric according to UV-Vis spectra. Antibacterial activity of the Ag/cotton samples shows that obvious bacteriostasis loops are observed around the samples with the appearance of both Gram-negative bacterium Escherichia coli (E. coli) and Gram-positive bacterium Bacillus subtilis (B. subtilis). The average diameter of the bacteriostasis loops against both E. coli and B. subtilis becomes larger with an increasing silver loading amount.This work provides a universal, fast, simple, and environmentally-friendly cold plasma method for synthesizing Ag NPs on heat-sensitive materials at atmospheric pressure.