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Archaeometric studies on mosaics often concentrate only on glass tesserae, while comprehensive studies including both stone and glass tesserae are scarce; however, both types of tesserae can sometimes bring relevant data to elaborate archaeological knowledge on a studied mosaic. In this paper, a representative set of tesserae from a large polychrome Roman mosaic retrieved in Barcelona (NE Spain) is investigated using various methods. Most of the techniques were directly applied on samples prepared as petrographic thin sections (including polarized-light, cathodoluminescence and electron microscopies, and synchrotron through-the-substrate μX-ray diffraction). The results indicate that, from the ten sampled stone tesserae, there are (i) seven limestones, one of them identified as Alveolina limestone (early Eocene) from the southern Pyrenees (ii) two sandstones from Barcelona's Montjuïc hill (Miocene) and, (iii) a Carrara white marble from the Apuan Alps (Italy). The profuse presence of tesserae of both local and imported materials with well-known uses in architecture, epigraphy, and sculpture could imply that tesserae were a by-product of their main use. Two different production technologies were identified for the three sampled glass tesserae. The concurrent use of antimony- and tin-based opacifiers is in agreement with the accepted archaeological chronology of the mosaic (4th century AD). [ABSTRACT FROM AUTHOR]