The electrospray deposition method has been used for preparation of catalyst layers for proton exchange membrane fuel cells (PEMFC) on Nation membrane. Deposition of Pt/C + ionomer suspensions on Nation 212 gives rise to layers with a globular morphology, in contrast with the dendritic growth observed for the same layers when deposited on the gas diffusion layer, GDL (microporous carbon black layer on carbon cloth) or on metallic Al foils. Such a change is discussed in the light of the influence of the Nation substrate on the electrospray deposition process. Nation, which is a proton conductor and electronic insulator, gives rise to the discharge of particles through proton release and transport towards the counter electrode, compared with the direct electron transfer that takes place when depositing on an electronic conductor. There is also a change in the electric field distribution in the needle to counter-electrode gap due to the presence of Nation, which may alter conditions for the electrospray effect. If discharging of particles is slow enough, for instances with a low membrane protonic conductivity. the Nation substrate may be charged positively yielding a change in the electric field profile and, with it, in the properties of the film. Single cell characterization is carried out with Nation 212 membranes catalyzed by electrospray on the cathode side. It is shown that the internal resistance of the cell decreases with on-membrane deposited cathodic catalyst layers, with respect to the same layers deposited on GDL, giving rise to a considerable improvement in cell performance. The lower internal resistance is due to higher proton conductivity at the catalyst layer-membrane interface resulting from on-membrane deposition. On the other hand, electroactive area and catalyst utilization appear little modified by on-membrane deposition, compared with on-GDL deposition. (C) 2010 Elsevier B.V. All rights reserved.