A comprehensive understanding of the behaviour of the heterogenous layers within the paint stratigraphies in historical paintings is crucial to evaluate their long term stability. We aim to refine nanoindentation as a new tool to investigate the mechanical behaviour of historical oil paints, by adapting the probes and the protocol already used in biomechanical research on soft tissues. The depth-controlled indentation profile performed with a spherical probe provides an evaluation of the non-linear viscoelastic behaviour of the individual layers in paint at local scale. The technique is non-destructive and guarantees the integrity of the surface after indentation. The mapping of elasticity demonstrates the properties’ heterogeneity of the composite material within the paint layers, as well as between the individual layers and their interfaces.