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The Hebrew Scroll, catalogued as Magliabekian Manuscript III 43 and belonging to the National Central Library of Florence (BNCF), is a membranous richly decorated scroll, with colorful depictions of sacred sites through the Holy Land to Lebanon along with handwritten texts in Hebrew and notes in Italian. Despite the fact that the manuscript was originally catalogued as an “object of no artistic or scientific value”, recent paleographic studies dated it to the XIV century and highlighted it as the oldest scroll still available, depicting holy places from Egypt to Lebanon. Nevertheless, precise dating, authorship, and the interpretation of its original function are still uncertain. A suite of complementary techniques was used, including photographic documentation in visible (VIS) light in diffuse light, grazing light, and transillumination, luminescence induced by ultraviolet (UV) radiation, imaging spectroscopy (IS), Macro Area X-ray Fluorescence (MA-XRF), and spot analyses such as fiber-optic reflectance spectroscopy (FORS) in the UV, VIS, and near-infrared (NIR) regions, Fourier transform infrared spectroscopy (FT-IR) in external reflectance mode (ER), and micro-Raman spectroscopy. The results of the non-invasive diagnostic campaign enabled the identification of several constituting materials (parchment, pigments, binder, and inks). The identified materials were consistent with the proposed dating and geographical manufacturing area of the artefact. Full article