Nanoparticles (NPs) are heavily used in biomedical, industrial, and commercial applications due to the benefits associated with the specific physical and chemical properties of both the bulk and the nanoscale material. The antimicrobial activity of NPs is widely recognized, but the mechanisms of their underlying toxicity remain unclear despite repeated attempts to establish a structure-function relationship between their physicochemical properties and their interactions with biological systems. [1,2] NP uptake in mammalian cells is generally considered to be an active process, mediated by endocytosis. Indeed, transport across the cell membrane and intracellular accumulation dictates the nanoparticle fate and cytotoxicity. [3] The critical size for NPs non-disruptively (passively) crossing cellular membranes is below 10 nm, irrespective of surface functionalization, [4-7] Figure 1. For this reason, there is a slowly forming consensus that smaller NPs bear greater toxicity than larger ones. [8,9] Similarly, the antibacterial properties of small NPs have It is commonly accepted that nanoparticles (NPs) can kill bacteria; however, the mechanism of antimicrobial action remains obscure for large NPs that cannot translocate the bacterial cell wall. It is demonstrated that the increase in membrane tension caused by the adsorption of NPs is responsible for mechanical deformation, leading to cell rupture and death. A biophysical model of the NP-membrane interactions is presented which suggests that adsorbed NPs cause membrane stretching and squeezing. This general pheno menon is demonstrated experimentally using both model membranes and Pseudomonas aeruginosa and Staphylococcus aureus, representing Gram-positive and Gram-negative bacteria. Hydrophilic and hydrophobic quasi-spherical and star-shaped gold (Au)NPs are synthesized to explore the antibacterial mechanism of non-translocating AuNPs. Direct observation of nanoparticle-induced membrane tension and squeezing is demonstrated using a custom-designed microfluidic device, which relieves contraction of the model membrane surface area and eventual lipid bilayer collapse. Quasi-spherical nanoparticles exhibit a greater bactericidal action due to a higher interactive affinity, resulting in greater membrane stretching and rupturing, corroborating the theoretical model. Electron microscopy techniques are used to characterize the NP-bacterial-membrane interactions. This combination of experimental and theoretical results confirm the proposed mechanism of membrane-tension-induced (mechanical) killing of bacterial cells by non-translocating NPs.