This paper presents the first systematic investigation of hand-painted magic lantern glass slides using multi-analytical techniques combined with a critical analysis of historical written sources of the painting materials and techniques used to produce them. The magic lantern was an optical instrument used from the seventeenth to the twentieth century that attained great success and impact on the entertainment industry, science, religion, and advertisement industry. The glass, colorants, and organic media of five magic lantern slides from the Museum of Natural History and Science of the University of Lisbon were studied. By means of energy-dispersive X-ray fluorescence spectrometry, the glass was characterized and the oxide quantification unveiled that the glass substrate was possibly produced between 1870 and 1930. Ultraviolet-Visible, Raman and Fourier transform infrared spectroscopies allowed the characterization of the colorants: Prussian blue, an anthraquinone red lake pigment of animal origin (such as cochineal), an unidentified organic yellow, and carbon black. The remaining colors were achieved through mixtures of the pure pigments. Infrared analysis detected a complex fingerprint in all colors, nevertheless, a terpenoid resin such as shellac was identified. Metal carboxylates were also detected, contributing to the assessment of the state of conservation of the paints.Heritage 2019, 2 2514 the subjects represented, which then included sequences of popular fiction, topographical themes, historical episodes, news items, educational and pedagogical subjects, as well as ludic scenes, generally combined with movement mechanisms [4,5]. During the transition to the twentieth-century, mainly lithographic and photographic slides were produced, especially for entertainment, pedagogical, and advertisement purposes [3,6].Research on magic lanterns and historical glass slides has been predominately centered on their role as a precursor to film and cinema. This research provides valuable information on the social, cultural, and economic relationships in the specific context of entertainment and the performing arts and has contributed to an understanding of the history and technical evolution of the apparatus and production processes [7]. Recently, there has been increased awareness that the magic lantern should be perceived as a medium and a cultural phenomenon representing a distinct screen practice [8]. The socio-political contexts in which magic lanterns were used has been a topic of historical media investigations [9]. Initiatives to develop appropriate systems for cataloguing and accessing this information are currently underway (e.g., in the European "A Million Pictures" project). However, despite this interest, no systematic information is currently available on the materials and techniques used in the production of magic lantern glass slides.To the extent of the authors' knowledge, there are only two papers concerning the characterization of magic lantern slides materials [10,11]. One of them studied the prese...