Rob Paulsen, who voices Donatello in this series, had previously voiced Raphael in Teenage Mutant Ninja Turtles (1987).
Before Rob Paulsen agreed to take the role of Donatello, he first wanted to ensure that his co-star Barry Gordon, who voiced Donatello in Teenage Mutant Ninja Turtles (1987), was okay with it.
In the second season, the device the turtles use to locate the ooze canisters is a modified original Nintendo Gameboy. Furthermore, the tone used when the device is activated is the same used when an original Gameboy is switched on.
In a few episodes, Michelangelo (Greg Cipes) can be heard saying "Go ninja! Go ninja! Go!" This is a reference to the Vanilla Ice song featured in Teenage Mutant Ninja Turtles II: The Secret of the Ooze (1991).
The Turtles strike a pose at the end of the title sequence (at the last reprise of "Heroes in a Half-Shell, Turtle Power" before the title card) is identical to the Turtles' pose on top of the building drawn on the cover of "Eastman and Laird's: Teenage Mutant Ninja Turtles" (vol.1) Issue #1 the original 1984 comic book from Mirage Studios.