The museum was inaugurated as the Etruscan Museum in 1870 and is one of the oldest archaeological museums in Italy. Its first location was at the Cenacolo di Fuligno and housed a collection of Etruscan, Greek, and Roman artifacts that came from the dispersal of the Medici and Lorraine collections previously kept at the Uffizi. In that same building, the Egyptian Museum had already been established in 1855. The spaces of the Cenacolo soon proved inadequate to accommodate the growing collections, and in 1880 the museum was moved to its current home in the Palazzo della Crocetta.
Today, the National Archaeological Museum of Florence includes the Etruscan Section, the Roman Section, the Greek Section, the Egyptian Section—also known as the Egyptian Museum (the second largest collection in Italy after Turin’s)—the Numismatic Section, and others.

Among the most important Etruscan artifacts are the Chimera, a votive bronze statue found in Arezzo in 1533 depicting a creature that is part lion, part goat, and part serpent. Other major works preserved here include the statues of the Arringatore (The Orator) and Minerva, the terracotta and travertine urns from Volterra, and Greek ceramic vases decorated with black-figure motifs.