A cultural heritage canvas from the early 19th century, painted by the Vaccaro brothers for the church of Niscemi, province of Caltanissetta, Sicily, was analyzed using Fourier transform (FT)-Raman, attenuated total reflectance-FT-infrared and surface enhanced Raman scattering (SERS) spectroscopy. The painting, still used in religious rites related to the Easter mass (‘la calata da tila’), depicts the scene of the Crucifixion and is executed in a scarce palette, with white, green and blue colors. Analysing vibrational data in conjunction with scanning electron microscopy and solid -state 13C-NMR signals of the linen threads, we were able to offer valuable insight into the painting technique, unknown prior to this study. SERS is usually employed in artwork diagnosis for the identification of organic lakes and dyes. Due to its sensitivity, SERS has been successfully applied for the detection of either organic painting materials (indigo) that are usually not resolved by conventional Raman spectroscopy or of inorganic pigments difficult to observe in the presence of highly fluorescent aged organic supports or binders. To the best of our knowledge, this is also the first report on the SERS investigation of flax used in linen from cultural heritage objects using Ag colloidal nanoparticles. Copyright © 2012 John Wiley & Sons, Ltd.