Natural oil bodies (OBs) from plant organs represent an important category of functional ingredients and materials in a variety of industrial sectors. Their applications are closely related to the membrane mechanical properties on a single droplet level, which remain difficult to determine. In this research, the mechanical properties of the membranes of OBs from soybean, sesame, and peanut were investigated in-situ by atomic force microscopy (AFM). Different regions of the force-deformation curves obtained during compression were analyzed to extract the stiffness K-b or Young's modulus of the OB membranes using Hooke's law, Reissner theory, and the elastic membrane theory. At higher strains (epsilon = 0.15-0.20), the elastic membrane theory breaks down. We propose an extension of the theory that includes a contribution to the force from interfacial tension based on the Gibbs energy, allowing effective determination of Young's modulus and interfacial tension of the OB membranes in the water environment simultaneously. The mechanical properties of the OBs of different sizes and species, as well as a comparison with other phospholipid membrane materials, are discussed and related to their membrane compositions and structures. It was found that the natural OBs are soft droplets but do not rupture and can fully recover following compressive strains as large as 0.3. The OBs with higher protein/oil ratio, have smaller size and stronger mechanical properties, and thus are more stable. The low interfacial tension due to the existence of phospholipid-protein membrane also contributes to the stability of the OBs. This is the first report measuring the mechanical properties of OB membranes in-situ directly. (C) 2020 Elsevier Inc. All rights reserved.