Twitch is seen with the other toys from Sunnyside Daycare in the Butterfly Room when the toys have their first donation. Mrs. Potato Head meets him and feels his biceps after helping her when she had fallen, much to Mr. Potato Head's jealosuly. Later that night, Twitch and the other of Lotso's workers enter a vending machine in Sunnyside Daycare playing using a See & Say toy.
Later on, he and the other toys find Buzz Lightyear out of the Caterpillar Room and seen capturing other toys under the Monkey's command. During confrontation with Lotso, Twitch and the other toys broke Chatter Telephone. Twitch and the rest of Lotso's workers retreated when Big Baby threw him into the dumpster.
When Sunnyside is restored, Twitch is waiting to be played at in the shelf by a toddler. He handles Chunk who is tossed by one of the toddlers.