Piazza Santo Spirito was born as an open-air theatre for the Augustinian monks to preach, and has been used also for non-religious purposes as well, such as shows and markets. The refectory is the oldest building of the monastery of Piazza Santo Spirito and was built in the second half of the 14th Century. The rectangular room has truss and two gothic-style windows on each of the long sides.

The frescoes (14th Century) decorating the room were severely damaged at the end of the 19th Century, when the refectory was used as a garage, and many details could not be recovered during restorations in 1941. In 1946 the antiquarian Salvatore Romano donated to the City of Florence his own collection of works of art: with them a new museum was established in the refectory of Santo Spirito.

In the interior of the refectory are still visible portions of a fresco ascribed to Andrea Orcagna (1360) and depicting the Crucifixion, the Last Supper (almost completely lost) and two Saints. In the room are collected several Sculptures: among them are ancient Roman figures, and sculptures by Tino da Camaino (Adorating Angel), from the school of Jacopo della Quercia, and two Bas-Reliefs by Donatello, once belonging to the Cathedral of Saint Antony in Padua. In the refectory are also displayed Frescoes and Furnitures from the 16th and 17th Centuries.