Abstract Zinc oxide (ZnO) is a promising semiconductor that is suitable for bioimaging applications due to its intrinsic defect fluorescence. However, ZnO generally suffers from poor photostability. We report room-temperature single-photon emission from optical defects found in ZnO nanoparticles (NPs) formed by ion implantation followed by thermal oxidation in a silica substrate. We conduct a thorough investigation into the photophysics of a particularly bright defect and identify other single emitters within the NPs. Photostability was observed when the NPs were removed from the growth substrate and taken up by skin cells for in vitro imaging.