After its foundation in 1895, the Museum had to wait several more years to assume a more concrete form. A preliminary list of 41 instruments acquired in the three preceding years and meant expressly for the Museum is registered in the 1897 Yearbook.

In 1900 the collection was probably shown to the public in the library located on Via Vittoria, as demonstrated by two photographs accompanying the description of the instruments written by academic Alberto Cametti in the Accademia's fifth Yearbook. During the three years following that first published list, the collection acquired 36 new pieces.

These instruments were combined with those used at the Liceo musicale (Conservatory), purchased for the Museum or donated by various collectors and Academics, as accurately indicated. Among the latter were the Count of San Martino, Branzoli and Berwin, as well as G. Basevi and Evan Gorga, who was also the former owner of the valuable collection now owned by the National Museum of Musical Instruments in Rome.

The Museum's combination of European and non-European folk instruments - about a third of the total collection - with ancient instruments from the classical tradition as well as more or less reliable reconstructions of the remote "ancestors" of modern instruments can be attributed to the taste for the primitive and exotic in vogue at the time. But it also demonstrates, in its subdivision according to the cataloguing criteria of the era, noteworthy scientific efforts and it reflects the desire to inform and communicate that initially inspired the creation of the collection. The mingling of classical and folk, European and non-European instruments establishes comparisons and connections that set this Museum significantly apart from its contemporaries, which tended to isolate the different traditions.