- Viewtiful Joe
- Jet Black
- Clowny - the Bianky of the Madow, Clownies are the most common enemy enountered in the game.
- Meta Rangers - These insect-like foes are comprised of Ranger Digi and Ranger Log.
- Killer Hands - The boss of Horror World, Killer Hands has many different weapons at the tips of his hands with which to fight. Appears to be based on Freddy Krueger and Edward Scissorhands.
- Queen Heinderella - the leader of the Madow. Queen Heinderella is the one responsible for the theft of the film, and the ultimate boss of the game.
In Double Trouble!, there are now 6 extra bars of VFX instead of 5. The Mach Speed and Zoom VFX powers present in the other Viewtiful Joe games have been removed. In addition, players collect V-Tapes instead of V-Films. The VFX bar also seems to drain slightly faster than in other games.
- Slow - Slows down the action to do more damage and begin lock-on. This is Joe's trademark power and has been in the series since the beginning, way back in the original Viewtiful Joe.
- Split - Allows the player to split the screen horizontally and can be used to drop objects on enemies as well as to jump through walls.
- Scratch - Allows the player to shake the screen around to make random pieces of debris fall on dazed enemies.
- Slide - Switches the bottom screen with the top. Slide is similar to Zoom from the first 2 games, but it also gives access to another power, Touch.
- Touch - Can be used to remove small, normally indestructible enemies, such as bats, from gameplay or to flip or turn switches in the background.
- The level Horror World is based on the Resident Evil series, featuring music and enemies from the games, and pictures of villains dressed as Jill, Carlos, Claire and Steve in the background.
- Following in the footsteps of Jadow and Gedow, the organization Joe and Jasmine square off against is called Madow.