The game is challenging because of the shapes of the plastic pieces and the narrow openings are barely larger than the pieces that they contain. In 2020, Hasbro introduced a new variation of the game called Operation Pet Scan in which players must remove objects from a dog's digestive tract. [5] In 2009, Hasbro released the first major update to Operation called Operation Sound FX. This is the first version of Operation to remove the buzzer and cards in place of a speaker. This meant that the game could announce the piece to be removed by the player by using an audio cue such as a "twang" sound for the rubber band and a busy tone for the cell phone. Players must press Sam's nose after successfully removing a piece; when all pieces have been removed and Sam's nose is pressed, the game plays a victory song. It also includes a standard game of Operation available by sliding the power button to the "lightning bolt" setting.

Licensed variations of Operation based on popular intellectual properties also exist, such as multiple versions of a Spider-Man-themed Operation with Spider-Man as the patient and a Fallout version in which the player must treat Vault-Tec's mascot Vault Boy and a "rads" alarm replaces Sam's nose buzzer.