Michelangelo Buonarroti (1475–1564) was a master anatomist as well as an artistic genius. He dissected cadavers numerous times and developed a profound understanding of human anatomy. From 1508 to 1512, Michelangelo painted the ceiling of the Sistine Chapel in Rome. His Sistine Chapel frescoes are considered one of the monumental achievements of Renaissance art. In the winter of 1511, Michelangelo entered the final stages of the Sistine Chapel project and painted 4 frescoes along the longitudinal apex of the vault, which completed a series of 9 central panels depicting scenes from the Book of Genesis. It is reported that Michelangelo concealed an image of the brain in the first of these last 4 panels, namely, the Creation of Adam. Here we present evidence that he concealed another neuronanatomic structure in the final panel of this series, the Separation of Light From Darkness, specifically a ventral view of the brainstem. The Separation of Light From Darkness is an important panel in the Sistine Chapel iconography because it depicts the beginning of Creation and is located directly above the altar. We propose that Michelangelo, a deeply religious man and an accomplished anatomist, intended to enhance the meaning of this iconographically critical panel and possibly document his anatomic accomplishments by concealing this sophisticated neuroanatomic rendering within the image of God.