Used in the nineteenth century for the display of ancient paintings from the collections of museum, the Galleria was later altered to house several very important sculptures by Michelangelo, thus creating a specific and unified itinerary that culminates in the centre of the Tribune where the statue of David stands.

The Galleria takes its name from the four large sculptures showing male nudes known as the Slaves or Prisoners. They were begun by Michelangelo for the tomb of Pope Julius II which was to have been built in the Basilica of St. Peter's in Rome. Left unutilised, after the death of Michelangelo they were presented to the Grand Duke of Tuscany, Cosimo I, who placed them in the Grotta del Buontalenti in the Boboli Gardens in Florence, where they remained up to 1909.

The fame of these four powerful statues is due above all to their unfinished state. Examples of Michelangelo's habitual working practice, referred to as non-finito, they are magnificent illustrations of the difficulty of the artist in carving out the figure from the block of marble, emblematic of the struggle of man to free the spirit from matter.

Also displayed here is the statue of St. Matthew, sculpted by Michelangelo in 1505-1506, the first and only one of a series of twelve Apostles commissioned from the artist for the Cathedral of Florence.

The Michelangelo collection is completed by the Pietà group, originating from the chapel of Palazzo Barberini in Palestrina, close to Rome. This was purchased by the Accademia in 1939, although expert critical opinion is now largely agreed in considering that it is not a work by Michelangelo.

In October 2006 several paintings by artists closely connected with Michelangelo, such as Granacci and Ridolfo del Ghirlandaio were placed in the Galleria dei Prigioni.