The Doni Tondo or Doni Madonna is the only finished panel painting by the mature Michelangelo to survive. Now in the Uffizi in Florence, Italy, and still in its original frame, the Doni Tondo commissioned by Agnolo Doni, probably to commemorate his marriage to Maddalena Strozzi, the daughter of a powerful Tuscan family. The painting is in the form of a tondo, meaning in Italian 'round', a shape which is frequently associated during the Renaissance with domestic ideas.

The Doni Tondo portrays the Holy Family (the child Jesus, Mary, and Joseph) in the foreground, along with John the Baptist in the middle-ground, and contains five nude male figures in the background. The inclusion of these nude figures has been interpreted in a variety of ways.

Vasari tells us that the work was commissioned by Agnolo Doni (portrait). The occasion may have been to celebrate his marriage in January 1504 to Maddelena Strozzi (portrait); the blazon of the Strozzi family with three crescent moons is carved in the frame.

Alternatively, it could have been commissioned to commemorate the birth of his daughter Maria in 1507. This last theory is supported by the noticeable influence that the Laocoon was unearthed in January 1506 in the presence of Michelangelo, and the similarities between the Doni Virgin and the Sybils in the Sistine Chapel frescoes, painted the following year.

In either case, Michelangelo completed it before started work in his frescoes in the Sistine Chapel ceiling, in 1508.

Mary is the most prominent figure in the composition, taking up much of the center of the image. She sits directly on the ground without a cushion between herself and the grass, to better communicate the theme of her relationship to the earth (?). Joseph is positioned higher in the image than Mary, although this is an unusual feature in compositions of the Holy Family. Mary is seated between his legs, as if he is protecting her, his great legs forming a kind of de facto throne. There is some debate as to whether Mary is receiving the Child from Joseph or vice versa. Saint John the Baptist, the patron saint of Florence, is very commonly included in Florentine works depicting the Madonna and Child. He is in the middle-ground of the painting, between the Holy Family and the background. The scene appears to be a rural one, with the Holy Family enjoying themselves on the grass and separated from the curiously (seemingly) unrelated group at the back by a low wall.

The painting is still in its original frame, one that Michelangelo might have influenced or helped design. The frame is ornately carved and rather unusual for the five heads it contains which protrude three-dimensionally into space. Similar to the nudes of the background, the meanings of these heads has been the subject of speculation. The frame also contains carvings of crescent moons, stars, vegetation, and lions' heads. These symbols are, perhaps, references to the Doni and Strozzi families, taken from each one's coat of arms. As depicted on the frame, "the moons are bound together with ribbons that interlock with the lions", possibly referring to the marriage of the two families.

There is a horizontal band, possibly a wall, separating the foreground and background. The background figures are five nudes, whose meaning and function are subject to much speculation and debate. Because they are much closer to us, the viewers, the Holy Family is much larger than the nudes in the background, a device to aid the illusion of deep space in a two-dimensional image. Behind Saint John the Baptist is a semi-circular ridge, against which the Sistine ignudi are leaning, or upon which they are sitting. This semi-circle reflects or mirrors the circular shape of the painting itself and acts as a foil to the vertical nature of the principal group (the Holy family). Mary and Joseph gaze at Christ, but none of the background nudes looks directly at him. The far background contains a mountainous landscape rendered in atmospheric perspective.