The church stands on the pre-existent oratory of the Servi di Maria (1235) which was built around the miraculous image of Our Lady of the Annunciation by seven young noblemen who decided to take monastic vows and give up worldly pleasures. As a further sacrifice, they later founded the Monastery of Monte Senario, above Fiesole.

Michelozzo built the First Cloister in the mid 15th century. The main body of the Church, started in 1440 by Michelozzo and Pagno Portigiani, was later altered by Alberti, who created the impressive Tribune that can be seen on the righthand side.

Three separate environments can be reached from the plain facade, decorated with the coats of arms of Pope Leo X dei Medici frescoed by the young Pontormo: the Chapel dei Pucci or of San Sebastiano on the right; the large Cloister dei Morti on the left, with frescoes also by Andrea del Sarto (the Madonna of the Bag); and the First Cloister or little Cloister dei Voti in the centre, frescoed throughout by masters of the Florentine Mannerist school of the early 16th century.

The interior of the church is dominated by the large tabernacle designed by Michelozzo to house the miraculous painting, itself now so frequently repainted that it is impossible to distinguish clearly. Florentine brides traditionally visit the shrine to leave their bouquets. Other chapels contain frescoes by Andrea del Castagno. The organ (1628) is the oldest in Florence and the second oldest in Italy. The circular tribune containing the high altar is unusual and is surrounded by chapels.

The church contains the tombs of the artists Baccio Bandinelli, Pietro Tacca and Giambologna. The cloister, known as the Chiostri dei Morti, contains the famous Madonna del Sacco by Andrea del Sarto. The Cappella di San Luca, which opens off it, has belonged to the Accademia delle Arti del Disegno since 1565. Many artists are buried in its vault, including Benvenuto Cellini, Pontormo, Franciabigio, Giovanni Montorsoli and Lorenzo Bartolini.