This iconic painting shows Dr. Tulp providing an anatomy lesson, as the forceps in his right hand lift a tendon from the partially dissected arm of a man who had been executed for armed robbery earlier that morning. Tulp looks toward the seven men gathered around the corpse as his left hand gestures to explain an anatomical point. The men, their white collars and fine clothes indicating their wealth, look in various directions. The three closest to the center lean forward as if watching Tulp's hands, while the two in the back look out toward the viewer. The two on the far left, depicted in profile, face toward Tulp but seem to be looking beyond him, outside the picture frame. A sense of dynamic movement and psychological complexity results, as no one returns Tulp's gaze or looks directly at the pale corpse. The umbra mortis, or shadow of death, fills the center of the canvas. The man's body, his genitals covered with a piece of white linen, evokes the iconography of Christ's death, though here, the body is forgotten, at the same time his dissected arm grimly conveys death's reality.

In this work, Rembrandt innovatively transformed group portraiture by dramatically focusing on the event in mid-action, rather than merely presenting a posed scene. As a result the work becomes a mise-en-scene, a kind of graphic documentary, and a masterful portrait.

The Amsterdam Guild of Surgeons commissioned the group portrait, Rembrandt's first important one in the city. The men attended the Guild's annual public dissection in 1632 at which Dr. Tulp, the City Anatomist, presided. In 17th century Holland, anatomical lessons were noted social events, accompanied by music, conversation, and food and wine, taking place in theater lecture rooms, and attended by those who could afford the entrance fee. The well-dressed appearance of these men, their white collars and fine gowns, testify to their social importance, and yet, they are presented as if both sensationalized and distracted, their humanity overshadowing their status.

Manet painted a copy of this painting in 1856 after studying it on a trip to the Netherlands, and the American realist Thomas Eakins was also influenced by it in painting his The Gross Clinic (1875). The executed man Kindt has also taken on a kind of later cultural life, referred to in W. G. Sebald's The Rings of Saturn (1999) and in Nina Siegal's The Anatomy Lesson (2014), which tells his story.