A running gag throughout the Ace Attorney series involves the protagonist and his assistant having a conversation about the difference between a ladder and a stepladder. First occurring in "Turnabout Samurai", the third case of Phoenix Wright: Ace Attorney, a stepladder has appeared in at least one location in each game to date (with the exception of The Great Ace Attorney: Adventures) for the sole purpose of continuing the tradition. The conversation generally goes as follows: after examination, the protagonist and one of their partner characters engage in a debate about the usage of the words "ladder" and "stepladder", and the difference between the two. Usually, one will accuse the other of being too absorbed in the details instead of looking at the big picture to realize that they both serve roughly the same function, to which the latter's response will often vary. Additionally, similar gags in regards to other items in the series have also been used, particularly in The Great Ace Attorney games, which primarily use a variant of the debate involving the difference between a shovel and a spade instead.

Within the various re-releases of the Ace Attorney games, an achievement is added that can be unlocked for engaging in every "ladder vs. stepladder" debate within the games featured in that collection. In the case of The Great Ace Attorney Chronicles, a second achievement for engaging in every "shovel vs. spade" debate within the two games is also included.

Regarding the difference between pixel art ("Classic" graphics) and digitally drawn art ("Modern" graphics) included in this collection, producer Shunsuke Nishida explained that he wanted to avoid using terms that would make one art style appear superior to the other like "remastered" or "refined", as he believed both had a significant amount of effort put into them and the only real difference between them when incorporated into the gameplay was the appearance.

Although the RE Engine was previously used to port the games in the Apollo Justice: Ace Attorney Trilogy, the Ace Attorney Investigations Collection instead used the Unity engine, as that was the engine used for the earlier mobile ports of both games, allowing for a cleaner porting process to the new collection.