Treatment of samples with plasmas in biomedical applications often occurs in ambient air. Admixing air into the discharge region may severely affect the formation and destruction of the generated oxidative species. Little is known about the effects of air diffusion on the spatial distribution of OH radicals and O atoms in the afterglow of atmospheric-pressure plasma jets. In our work, these effects are investigated by performing and comparing measurements in ambient air with measurements in a controlled argon atmosphere without the admixture of air, for an argon plasma jet. The spatial distribution of OH is detected by means of laser-induced fluorescence diagnostics (LIF), whereas two-photon laser-induced fluorescence (TALIF) is used for the detection of atomic O. The spatially resolved OH LIF and O TALIF show that, due to the air admixture effects, the reactive species are only concentrated in the vicinity of the central streamline of the afterglow of the jet, with a characteristic discharge diameter of ∼1.5 mm. It is shown that air diffusion has a key role in the recombination loss mechanisms of OH radicals and atomic O especially in the far afterglow region, starting up to ∼4 mm from the nozzle outlet at a low water/oxygen concentration. Furthermore, air diffusion enhances OH and O production in the core of the plasma. The higher density of active species in the discharge in ambient air is likely due to a higher electron density and a more effective electron impact dissociation of H 2 O and O 2 caused by the increasing electrical field, when the discharge is operated in ambient air.