In 1932, Calloway recorded the song for a Fleischer Studios Talkartoon short cartoon, also called Minnie the Moocher, starring Betty Boop and Bimbo. Calloway and his band provides most of the short's score, and appear in the short themselves in a live-action introduction. The thirty-second live-action segment is the earliest-known film footage of Calloway.

In the cartoon, Betty decides to run away from her harsh parents (to the tune of "Mean to Me"), and Bimbo comes with her. While walking away from home, Betty and Bimbo wind up in a spooky area, and hide in a hollow tree. A ghost walrus—whose gyrations were rotoscoped from footage of Calloway dancing—appears to them, and begins to sing "Minnie the Moocher", with many fellow ghosts following along. After singing the whole number, the ghosts chase Betty and Bimbo all the way back to Betty's home. While Betty is hiding under the covers of her bedsheets, her runaway note is torn up and the remaining letters read "Home Sweet Home".