The painting materials used by Lucio Fontana were investigated through the analysis of 28 Spatial Concept works created between the early 1950s and the artist's death in 1668. The pictorial media were analyzed by means of numerous techniques including Fourier transform infrared spectroscopy , pyrolysis-gas chromatography-mass spectrometry, scanning electron microscopy, and X-ray fluorescence. This study has provided a more complete understanding of the artist's technique, as ir was found that Lucio Fontana had already begun experimenting with new synthetic pictorial and binding media such as alkyds and polyvinyl acetates at the beginning of the 1950s, and polyesters during the early 1960s. Although he never abandoned the use of commercial oil paints, Fontana's application of these new materials continued in an infinitely varied search for innovative expressive effects. The characterization results explain the physical and mechanical properties of the paint layers, and may be correlated with the conservation state of the works analyzed. All the results have been compiled and are discussed together with the artist's practices and their evolution over time.