Understanding the interaction of graphene oxide (GO) with a lipid membrane is important for the development of tissue engineering and to advance graphene-based biology. In an effort to understand the GO-lipid membrane interaction, appropriate characterisation of GO structure was determined by using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and a field emission scanning electron microscope (FESEM). In this study, the lipids 1,2-dipalmitoyl-sn-glycero-3phosphocholine (DPPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were used to produce a lipid vesicle with a conventional gentle hydration method and observed under transmission electron microscopy (TEM). Lipid vesicle-GO interactions were also investigated using dynamic light scattering (DLS) (Malvern Zeta) and TEM by focusing on the effect of the surface charge interactions and localisation of GO on the surface of the vesicle membrane. It was observed that the surface charge of the vesicles increased as the GO nanoparticle concentration increased, but for the low saturation lipid the surface charge remained high as the nanoparticle concentration increased. The localisation and positioning of the GO nanoparticles in the lipid vesicles were confirmed with TEM analysis.