A combination of large-scale and micro-scale elemental imaging, yielding elemental distribution maps obtained by, respectively non-invasive macroscopic X-ray fluorescence (MA-XRF) and by secondary electron microscopy/energy dispersive X-ray analysis (SEM-EDX) and synchrotron radiation-based micro-XRF (SR m-XRF) imaging was employed to reorient and optimize the conservation strategy of van Eycks renowned Ghent Altarpiece. By exploiting the penetrative properties of X-rays together with the elemental specificity offered by XRF, it was possible to visualize the original paint layers by van Eyck hidden below the overpainted surface and to simultaneously assess their condition. The distribution of the high-energy Pb-L and Hg-L emission lines revealed the exact location of hidden paint losses, while Fe-K maps demonstrated how and where these lacunae were filled-up using an iron-containing material. The chemical maps nourished the scholarly debate on the overpaint removal with objective, chemical arguments, leading to the decision to remove all skillfully applied overpaints, hitherto interpreted as work by van Eyck. MA-XRF was also employed for monitoring the removal of the overpaint during the treatment phase. To gather complementary information on the in-depth layer build-up, SEM-EDX and SR m-XRF imaging was used on paint cross sections to record microscale elemental maps.Several methods for obtaining information about the distribution of pigments and other chemical constituents of artworks have been developed; [1] such methods are necessarily non-invasive and employ radiation in the infrared, visible, ultraviolet or X-ray range. Traditionally, the spectroscopic characterization of artworks was done on a point-by-point basis, an approach that has inherent limitations with respect to representativeness. The recent transformation of spectroscopic instrumentation into mobile scanning systems [2][3][4][5] that allow the examination of all locations on the surface of (flat) artworks, proved to have a significant added-value for the study of cultural heritage objects. These forms of (spectro)-chemical imaging allow the consideration of large and complex spectroscopic datasets as (hyperspectral) image stacks that are (more) easily understandable by non-chemists such as art conservators and art historians than extensive series of spectral data derived from individual points. Whereas subtle point-to-point variations in the spectral response may be hard to notice and even more difficult to interpret correctly, gradual compositional variations that become apparent in large-scale chemical or elemental maps are usually much easier to link to (in)visible features of the paint surface and/or its conservation state.Since its introduction by Alfeld et al., [2] mobile MA-XRF scanning has met with considerable attention and success, for example, by supplying new and pivotal arguments for authentication of high-profile works of art in cases in which more conventional analytical and/or imaging techniques led only to ambiguous in...