The significance of the Labyrinth in the myth of Theseus and the Minotaur lies in its representation of fear, conflict, and the journey to overcome challenges. The Labyrinth was designed by Daedalus to contain the Minotaur, a symbol of humanity's inner demons. Theseus’s successful navigation and subsequent defeat of the Minotaur signify triumph over adversity, emphasizing themes of heroism and self-discovery.