Allegedly built where Saint Ambrose would have stayed when in Florence in 393, the church dates back to 998 as a chapel of a nunnery built in honour of the saint. It was rebuilt in the 19th century but still has an original open timber roof.

The church contains numerous frescos, altarpieces, and other artwork attributed to Andrea Orcagna, Agnolo Gaddi, Niccolò Gerini, Lorenzo di Bicci, Masaccio, Filippo Lippi, Sandro Botticelli, Alesso Baldovinetti, Mino da Fiesole, Cosimo Rosselli, Fra Bartolomeo, and many other artists.

Several important artists are buried in the church, including Francesco Granacci, an Italian painter of the Renaissance and lifelong friend of Michelangelo Buonarroti, the painter and sculptor Verrocchio, the architect Cronaca, and the sculptor Mino da Fiesole.