Cenacolo di Santo Spirito, Gothic refectory museum in Santo Spirito, Italy

The Cenacolo di Santo Spirito is a former dining hall of an Augustinian monastery located at Piazza Santo Spirito in Florence. Its eastern wall displays a large fresco showing the Last Supper and Crucifixion, which dominates the space with religious imagery.

The refectory was part of the Augustinian convent and survived Filippo Brunelleschi's 15th-century rebuilding. In the late 19th century it served as a tram depot; opening a wall onto the piazza then destroyed Andrea Orcagna's Last Supper. Above its remains is a large Crucifixion painted between 1360 and 1365. From 1894 to 1928 the sculptor Raffaello Romanelli used the hall as his studio.

The hall holds the Salvatore Romano Foundation's collection of sculpture from the 11th to 15th centuries, donated to the city by the antiquarian Salvatore Romano. Highlights are two bas-reliefs attributed to Donatello from Padua, Jacopo della Quercia's Madonna and Child, and two marbles by Tino da Camaino from 1320–1322. Detached frescoes and 16th–17th-century furniture are also displayed.

The museum is located at Piazza Santo Spirito 29 and is easily reached on foot from central Florence. The entrance is straightforward, and the gothic dining hall represents the only surviving gothic section of the former Augustinian monastery.

The wall facing Piazza Santo Spirito was opened in the late 19th century, when the hall was used as a tram depot; this destroyed Andrea Orcagna's Last Supper beyond repair. Above the fragmentary fresco survives a large Crucifixion painted between 1360 and 1365.