Director: Masakazu Hashimoto

Based on the popular and award-winning Professor Layton Nintendo DS puzzle game series, this movie is set to take place directly after the events of “Professor Layton and the Specter's Flute”, in which Layton's apprentice, Luke, is investigating the concept of immortality.

Professor Layton receives a letter from the famed opera singer, Jenice Kaitly, inviting them to see her newest performance. She also asks Layton, Luke, and Remi for assistance in a mystery, saying her friend Melina went missing one year ago but returned later as a seven year old girl saying she had found the secret of eternal life. Naturally, the case fascinates Layton and his apprentices, so they head to the Crown Petone opera house. When they arrive, something is very strange: the pianist, Ozro Vislar talks of a game, and the winner will win the "eternal life." It is up to Layton, Luke, and Remi to solve the mystery of the Eternal Diva before it is too late.