The Department is heir to the Physics Institute founded in 1914 and moved to the Arcetri centre in 1921. Leading figures in the Department's first stage of life were the physics professors Antonio Roiti and, after him, Antonio Garbasso. This Department of the Florence University was dynamically active in research and in modernising the orientation of study by introducing new teaching methods, as exemplified by the Department of Mathematical Physics directed by Enrico Fermi. During World War II, research activity was severely restricted, to be resumed again in the 1950s. The Department, which has recently moved to the campus at Sesto Fiorentino, is now in the process of reorganising its collections of instruments and bibliographical material.

The physics collection contains instruments constructed in the late 19th and early 20th centuries: galvanometers, electrometers, standard resistors, compasses, current balances, sets of weights, microscopes, division scales, a magnetometer and a harmonium. Some of these instruments, including prototypes constructed by the institute iself, constitute rare examples of their typologies.