Today part of the Accademia Gallery, the Museum preserves a precious antique collection of musical instruments that can be dated between 1568 and the 20th century, the original nucleus of which originates from the acquisitions made by the Medici court. The magnificent court balls, concerts and the first performances of opera, born in Florence in 1600 in the ambit of the "Camerata de’ Bardi" (whose members also included Vincenzo Galilei, Galileo’s father), aroused particular attention in the Medici princes in their purchase of instruments of increased technical perfection, patronising the activity of lutists and organ makers. One example is the famed Bartolomeo Cristofori who was summoned to Florence where, in 1698, he built the first piano of which, however, only Cristofori’s description remains. The Museum’s most important instruments feature violins, violas and cellos by Antonio Stradivari, a cello by Amati, an upright piano by Domenico Del Mela, a spinet by Floriani; the wind instruments include the Giorgi flute, the Briccialdi flute and a "bimbonifono", that is to say a trombone equipped with a mechanism that also allowed other, deeper mouthpiece instruments to perform particular virtuoso passages such as legatos, trills, etc., invented by Giovacchino Bimboni between 1849 and 1870. Enriched in time by new acquisitions, in 1863 the collection was transferred from the Pitti Palace to the "L. Cherubini" Music Conservatory where it was inaugurated in 1926.

The Museum exhibits about fifty musical instruments from the private collections of the Medici and Lorraine grand dukes of Tuscany, collected between the second half of the 17th century and the first half of the 19th century, and later transferred to the Cherubini Conservatory of Florence which, since 1996, has given them on free loan to the Accademia Gallery. These include outstanding pieces such as: a tenor viola by Antonio Stradivari, the cello that belonged to the same quintet made in 1690 for Grand Prince Ferdinando, a Stradivarius violin from 1716, and a cello by Niccolò Amati from 1650.

The Museum also exhibits two instruments built by inventor of the piano Bartolomeo Cristofori, and the oldest upright piano conserved today. The instruments are presented alongside paintings depicting scenes of musical life at the Medici court. A series of computers permits visitors to hear the sound of all the instruments exhibited, as well as to have information on the musical culture of grand-ducal Florence. The museum is part of the Accademia Gallery.