The two paintings Infant Bacchanals (Museo Nazionale d’Arte Antica, Palazzo Barberini, Rome, Italy) executed by Nicolas Poussin (Les Andelys, 1594–Rome, 1665) in around 1626 are thought to have been painted “a guazzo”, which means either with a glue or with an egg binding medium. To date, this has never been confirmed through analysis. Dual-beam time-of-flight secondary ion mass spectrometry (TOF-SIMS), using a bismuth cluster liquid metal ion gun and an argon gas cluster ion beam, allows the mapping of organic and inorganic matter on paintings cross sections, with the possibility to acquire submicrometer-resolution mass spectrometry images of the sample, together with high mass resolution using a delayed extraction of secondary ions. The surfaces of cross sections from both paintings were prepared beforehand either by polishing or by microtome cutting and then cleaned with the gas cluster ion beam directly inside the vacuum chamber of the instrument. The nature of the binders in the two paintings was investigated by TOF-SIMS analyses. By considering the uneven physical properties of the heterogeneous analyzed surfaces, several high-resolution images were recorded with different instrument settings. The detection of lipids seems to point toward an oil-containing medium, rather than a glue-binding medium. An emulsion made of oil and glue is another hypothesis to be explored to better understand the artist’s working methods in his early career.