The mechanical behavior of nanobubbles represents their physical essence and has been thought to be closely related to their mysteriously long lifetimes. However, it is difficult to measure the mechanical properties of nanobubbles by conventional atomic force microscopy (AFM). In this paper, nanobubbles were investigated via a novel AFM imaging mode, PeakForce Quantitative Nano-Mechanics (PF-QNM), at the interface of water and highly oriented pyrolytic graphite (HOPG). High resolution images of the nanobubbles in true-contact were achieved by PF-QNM and compared with those obtained by tapping mode AFM (TM-AFM) in the same area. From the force curves simultaneously captured during the PF-QNM imaging processes, the stiffness of the nanobubbles was derived and mapped, ranging usually from 60 to 120 pN nm⁻¹, indicating that the gas–water interface of nanobubbles has similar mechanical properties to those of microbubbles. Interestingly, a size dependence of the stiffness was found and the small nanobubbles had a higher stiffness.