Professor Hershel Layton—renowned archaeologist, puzzle-solver, and extraordinary gentleman—and his young aspiring apprentice Luke Triton receive a mysterious letter from his former student Janice Quatlane containing tickets to an opera at the Crown Petone. They soon discover that it is no ordinary opera, but rather a devious game for the legendary Fountain of Youth—the giver of eternal life. However, this only scratches the surface of an unfolding mystery that the pair must solve together in order to survive.

Despite being faced with complex puzzles and fervent competition, Professor Layton and his trusted assistant Luke never forget to uphold the standards of true gentlemen... and a true gentleman never leaves a puzzle unsolved.