Pop'n Twinbee has gameplay similar to Detana!! Twinbee, the second coin-op game in the series, but with a few notable additions and changes. Like in previous TwinBee, up to two players can play simultaneously, Player 1 controls TwinBee (the blue aircraft), while Player 2 controls WinBee (the pink aircraft) When the player begins the game, they'll be asked to enter their name and choose between three types of companion ships (whether their companions will follow their ship, engulf them, or surround them). The player's objective is to maneuver their spacecraft through a series of seven stages and defeat every enemy who gets in the way, facing a boss at the end of each stage. Unlike previous games, instead of extra lives, the player has a life meter that determines how much damage the player's ship can take before losing. When the player loses, they will be given a chance to restart at the stage where they left off, but only limited chances are given to continue.