The collection comprises about fifty instruments, dating from the late 17th to early 19th centuries, formerly belonging to members of the Medici and Lorraine families, which later passed to the Cherubini Conservatory of Florence. Since 1966 they have been deposited on loan to the Accademia Gallery where they are currently displayed.

Among the most interesting instruments on display are a tenor viola by Antonio Stradivari, a violoncello which formed part of the same quintet of strings made in 1690 for Grand Prince Ferdinando, a violin of 1716 by Stradivari and a cello of 1650 by Niccolò Amati. The Museum also has two rare instruments by Bartolomeo Cristofori, the inventor of the piano, and the oldest vertical piano in existence. The instruments are exhibited with paintings representing musicians at the Medici court. Computers enable visitors to hear the sound of all the instruments on view besides providing information on the history of music in Florence at the time of the grand-dukes.

The Museum forms part of the Accademia Gallery.