Established at the initiative of Francesco Marucelli as a public library for the less affluent citizens, it opened to the public on September 18, 1752, on the same premises it occupies today. In 1783 the collection of drawings and prints was inaugurated. The Library’s wealth of books was considerably increased subsequent to the suppression of the ecclesiastical bodies, between the 18th century and the early years of the 19th. Other important sources were added during the first years of the Kingdom of Italy. Since 1910, thanks to the law instituting the compulsory deposit of printed materials, the works printed in the Province of Florence are deposited here. Today the Library possesses an imposing heritage made up of over 500,000 volumes (including incunabula and sixteenth-century editions), manuscripts and unbound papers (autograph works and correspondence of literary, historic, artistic and scientific interest), periodicals and opera libretti, in addition to an important collection of prints and drawings (15th to 19th centuries). Noteworthy among the autograph works are the papers of Francesco Redi, Giacinto Cestoni and some of the letters of Antonio Vallisneri.

The Library also contains the "Luigi Beuf" Italian Cryptogamic Herbarium. The Herbarium consists of a collection of dried cryptogams (around 3,000 samples) glued to removable sheets. This work, kept in the Manuscripts and Rare Items Room, was produced in thirty instalments by the Luigi Beuf European Library of Genoa, between 1858 and 1867.

Especially important from the scientific viewpoint is a pair of globes (celestial and terrestrial) from 1699, the work of the Venetian cosmographer Vincenzo Coronelli, purchased from the Bandini Library in 1755.