Narrative: Detective / Mystery
Published by: Nintendo of America Inc
Developed by: Level-5 Inc
Platform: Nintendo DS (NDS)
The game includes, only in the American and Japanese versions, also an RPG known as "Life of London" which can be unlocked after the conclusion of the puzzles.
After receiving a mysterious letter asking for his help, the professor is invited by an old friend to go to Misthallery, a city on the water characterized by frequent fog and where strange events are happening. In fact, in the village it seems that a huge spectrum is destroying all the buildings piece by piece, appearing only during the fog and accompanied by a mysterious music. In this adventure, the first chronologically speaking, Professor Layton will leave only from his university office and then meet during the unfolding of Emmy events, with which he will travel to Misthallery. In the plots of the plot, this being a prequel, the first meeting between the professor and Luke Triton is also told.
- Professor Hershel Layton: A professor of Archeology at the Gressenheller University. He became a professor at the age of 27. He is 35 years old and is a great fan of puzzles and mysteries.
- Luke Triton: Son of Clark Triton. He has a spectacular ability to understand animals. He will also become Professor Layton's apprentice.
- Emmy Altava: Assistant to Professor Layton, nominated by the president of the Gressenheller university. She considers herself a heroine, and loves taking photos, always carrying her camera with her.
- Arianna Barde:Luke's friend. All the inhabitants think that she is a witch because she isolated herself from everyone for her deadly disease. It will heal thanks to the sacrifice of Loosha.
- Inspector Clamp Grosky: He's a Scotland Yard police inspector (replaced by Inspector Chelmey), he has a burning passion for the mysteries that require his experience. He is also very athletic and energetic.
- Oscar: It's a cat that gathers all the unsolved puzzles to help Layton solve them.
- Rector Stone: Dean of the Gressenheller University, and therefore is the leader of Layton. Contrary to Layton's wishes, he hired Emmy as his assistant.
- Clark Triton: Mayor of Misthallery and father of Luke. He's also an old friend of Layton's. It was he who requested Professor Layton's help to solve the mystery.
- Loosha: A giant marine animal and a close friend of Arianna and Tony. He sacrificed his life to heal Arianna from his illness.
- Tony Barde: Arianna's younger brother.
- Jean Descole: Layton's worst enemy. Very little is known about him and it is not known how he and the professor met.
- Roland Noble: Triton family butler. He has a strong connection with Luke.
- Henry: Mysterious gardener of the Barde family. Because of its terrifying face, the inhabitants fear it. He is very protective of Arianna and hates visitors. Later it turns out to be Tony in disguise.
- Jakes: Misthallery Police Chief. It is estimated by all inhabitants. Later it will be discovered that he was an accomplice of Descole.
- Toppy: Luke's little mouse. Turn to Misthallery to find clues and tell her friend.
- Crow Torvo: The demon that attacks Layton, Luke and Emmy, chasing them. In a sense it will help the professor.
- Crow: Head of the Torvi Crows band. It will help Layton build the catapult to stop the spectrum.
- Scalmo: It is used to move quickly with the boat, it is a character that uses the boat even in the absence of water.