The upsurge in the number of thin film products has encouraged studies into every aspect of their fabrication and application. An additional source of industrial interest is the laser ablation of thin films to generate nanoparticles. This technique offers advantages over other fabrication methods, as no chemical pre-cursers are required, thereby giving rise to a pure product. The main disadvantage lies in the difficulty with controlling the size of the nanoparticles. This study aims to clarify the influence of the microstructure of a thin film on its optical properties and also to establish the size relationship between the film grain and the nanoparticles generated during laser ablation. A comprehensive sample set of Gold (Au) films with different grain sizes was achieved using different deposition rates, temperatures, film thicknesses (<100 nm) and substrates: Silica, Quartz and Sapphire. The microstructure of each film was analyzed using Atomic Force Microscopy (AFM). Single femtosecond laser pulses, above the ablation threshold fluence of each film, were applied to generate nanoparticles. Scanning Electron Microscopy (SEM) was used to image the re-deposited nanoparticles, from which the nanoparticle size distribution was established. Results confirm that the film microstructure is directly linked to the nanoparticles generated during laser ablation. (C) 2012 Elsevier B. V. All rights reserved.