In Presentation of the Virgin in the Temple, Giotto portrays a young Mary entering the Temple of Jerusalem, where she would be consecrated and educated in her role as Mother of God. Her parents, Joachim and Anne, at the bottom of the stairs, guide the three-year-old Mary to the waiting arms of the priests. The New Testament does not relate this story, but several of the apocrypha do. A corresponding feast in celebration of this day exists, as well as a religious order, the Presentation Sisters. In this remarkable fourteenth-century fresco, Giotto displays a grasp of perspective and compositional techniques far ahead of his time, although perhaps still awkward to the modern eye.

The colors have remained vibrant over the years, allowing the deep reds on Anne's dress and the priest's robes to stand out and balance each other. Between them, the dress of the pure, virginal Mary remains the brightest white in the piece. The walls of the temple create a clear horizontal delineation between the sky and the ground, while the thin columns create a vertical barrier, creating four quadrants. In addition, the line of the staircase and the pointing hand of the figure on the right lead the eye to Mary, and each person's eyes focus on Mary, leaving the viewer in little doubt who the most important person in the painting may be. Giotto shows a truly remarkable level of skill in this fresco and in the details of the faces, the embellishments on the temple, and the folds of the clothing.