Imaging and punctual analysis to support the bio-cleaning, through bacteria-based system, of the late 16th canvas painting “Coronation of the Virgin with the Saints Ambrose and Jerome”, attributed to the Tuscany-Umbria area.

The aim of this work is to combine non-invasive imaging with chemical characterization analyses to study original and restoration materials of a late 16th-century painting on a canvas representing the “Coronation of the Virgin with the Saints Ambrose and Jerome”, preserved in the Diocesan archive of Orte, a town in the district of Viterbo (Italy). The diagnostic campaign was addressed to support the restoration activities and the choice of the most suitable cleaning operations. (2) Methods. Both traditional analytical techniques and innovative multispectral imaging were applied to solve the diagnostic issues and best address the restoration of the painting. Specifically, hypercolorimetric multispectral imaging (HMI), X-ray fluorescence spectroscopy (XRF), Fourier transform infrared spectroscopy (FT-IR), optical microscopy, and gas chromatography coupled with mass spectrometry (GC-MS) were combined to obtain information on the general conservation state of the artwork and the characterization of pigments, organic binders, and superimposed materials, these last being particularly important to identify ancient and not-documented restoration intervention, enabling the correct choice of the most suitable and effective cleaning intervention. (3) Results. Multispectral data allowed us to differentiate and map original materials through infrared and ultraviolet false color images and spectral reflectance-based similarity maps, suggesting pigment attribution and focusing point analysis for characterization. This approach was particularly successful to identify and locate the presence of unaltered smalt blue in the first painting coat, which had been covered with other pigments, and to suggest the use of organic dye in mixtures with cinnabar and ochres. Spectroscopic and chromatographic techniques enabled us to identify the painting palette and confirm the use of oil-based binder for the pigments and characterize the altered top layers, made with a natural resin and an animal glue. (4) Conclusions. The characterization of the artwork’s materials was essential to select the most suitable methods and materials for the bio-cleaning, based on bacteria, experimented with during the restoration activities.