This time around, Luigi must travel through five different mansions, each with their own theme, solve puzzles along the way and defeat Possessors and retrieve Dark Moon pieces. Also, Luigi gains the Dark-Light Device, which can make any invisible object visible once again. Simply shining the flashlight on ghosts no longer stuns them, so Luigi must activate the Strobulb function on his new and improved vacuum, allowing Luigi to stun and vacuum up any ghosts that stand in his way. There is now an upgrade to the Poltergust 3000, the Poltergust 5000. The ghosts spend a large amount of time hanging out with Luigi and co., whilst Luigi also comes to own a friendly ghost pup, Polterpup. Once he is defeated again, Luigi frees his brother, helps the Toads to complete the Dark Moon and restore the ghosts of Evershade Valley back to their peaceful selves. Afterwards, he is dragged into King Boo's Illusion to fight the king of illusions himself.

With his brother on the line, Luigi eventually travels to the Paranormal Dimension to collect the final Dark Moon piece. Gadd figure out that King Boo is behind the cause of the events, and has captured Mario once more. Luigi will travel through 5 different mansions in search of the Dark Moon pieces, fighting swarms of Boos and ghosts and solving puzzles along his journey. He then tasks Luigi to help him capture ghosts once more, equipped with the newly-built Poltergust 5000, Dark-Light Device and Strobulb. However, King Boo appears and destroys the Dark Moon, causing the ghosts to become hostile and forcing the professor to evacuate.

In Evershade Valley, Professor Elvin Gadd is working on some new inventions with some friendly ghosts.

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