The present study focuses on an icon known as the Panagia Valieriotissa at the Church of Saint Nicholas in Palodeia, Cyprus. The icon is unusual as it contains two depictions of the Virgin Mary and Child. The main aim of this study is to analyze the material characteristics of the icon and the relation of the two panels in order to determine if the icon belongs to a distinct category of paintings known as “composite icons” – icons consisting of two separate panels, one inserted into another. Icons of this unusual type, first described by Panayotis Vocotopoulos, appear to be quite numerous on Cyprus and date mostly to the sixteenth century. Results show that the icon can be included within this group, consequently enriching our knowledge on the materials, technologies, and techniques used in the creation of composite icons. Driven by art historical inquiries, the study’s methodological approach was based on the integration of digital imaging techniques with non-invasive and non-destructive material analyses pursued by the Andreas Pittas Art Characterization Laboratories (APAC Labs) of the Cyprus Institute. Spectroscopic techniques, digital microscopy and advanced digital imaging methods produced a broad range of data which were analyzed and interpreted from both an art-historical and a conservation perspective towards a better understanding of the materiality of the Palodeia icon and its subsequent history and use. The imaging and spectroscopic methods used provided new information on the materials and techniques used by the painters as well as on later undocumented episodes of restoration or repainting. Overall, the results showed that the icon indeed consists of two separate icons – a larger panel and a smaller, re-used icon inserted into the former. Moreover, some important distinctive features of the inset icon, invisible to the naked eye, reveal its possible previous use and dating. The study brought to light the great potential of non-invasive methods in the revealing and analysis of unknown aspects of composite icons, thus shedding light on this fascinating phenomenon, but it can also interest a broader group of researchers into the exciting new possibilities as well as the limitations of non-invasive methods for the study of works of art and heritage objects.