Scanning electron microscopy in combination with transmission and reflectivity spectroscopy have been employed to characterize the structural, morphological and optical properties of thin gold film deposited on polystyrene colloidal crystal. Surface-enhanced Raman (SER) spectra of the probe Rhodamine 6G molecules adsorbed on the substrate have been also recorded by using the 514 and 633 nm laser lines for excitation in order to evaluate the performance of the gold nanostructured film as SERS-active substrate. Interesting correlations between the optical response, nanometer-scale morphology and SERS activity of the nanostructured film have been found. (C) 2005 Elsevier B.V. All rights reserved.