Gotham Girls is one of the few Flash animation web series made by a major mainstream cartoon publisher, using Flash and vector graphics so the animation remains undistorted at any resolution.

The series reused the same professional voice cast as Batman: The Animated Series and The New Batman Adventures, including Arleen Sorkin, Diane Pershing, Adrienne Barbeau, Tara Strong, and Bob Hastings reprising their roles.

Because the show used Flash symbols only drawn within the intended viewing frame, animated elements like a light beam on Harley Quinn in the first episode continue off-screen while her body is only rendered within the visible dimensions.

Each episode included a mini-game or puzzle to play while the clip downloaded, plus an interactive feature letting viewers help a character make a decision.

While the first two seasons were mostly comedic, the third season became more serious, dropped the magician Zatanna, added Detective Renee Montoya, and tied directly into Batman & Mr. Freeze: SubZero, forming a continuous half-hour whodunit dealing with police corruption and Poison Ivy's environmental crusade.

The series finale, 'Cold Hands, Cold Heart' (2002), was dedicated to the memory of Hilary J. Bader, a script writer for numerous DC Animated series, who died on November 7, 2002, of breast cancer.

The complete Gotham Girls series was later included as a bonus feature on the DVD release of the Birds of Prey television series in 2008, presented in linear fashion without the original interactive decision-making option.

DC Comics produced a five-issue Gotham Girls comic book miniseries in 2003 written by Paul Storrie, set in the DC animated universe, which was followed by a three-issue miniseries called Batman: Harley and Ivy.