* ''Webcomic/PennyArcade'' complained about having to sort out "some jackhole's insane chicken scratches." [[http://professorlayton3walkthrough.blogspot.com/2009/01/puzzle017.html Puzzle 17]] in ''Unwound Future'' has you do just that.
* When Luke, Layton, and [[spoiler:Don Paolo]] are escaping through an underground secret passage, which is too dark to see anything.
--> '''Luke:''' Ow! What did I just walk into?
--> [[spoiler:'''Don Paolo:''']] I don't know, maybe a wall?
* In the flashback when [[spoiler: Don Paolo finds out Claire is in love with Hershel]], he runs away crying. ''Into a river.'' '''''[[SuperNotDrowningSkills And he doesn't notice.]]'''''
* Really, [[spoiler:Don Paolo]] had a good deal of funny moments in this game. Another good one would be when [[spoiler:he {{lampshade}}s Luke making an overly obvious remark by ''outright calling him'' CaptainObvious.]]
* They dedicated a cutscene to revealing who Granny has sent to take over her job. [[spoiler: Surprisingly, it's not the bee we've been dealing with.]] It just pops out of nowhere and it's an actual cutscene...
** Especially [[spoiler:Puzzlette coming out of nowhere and swatting Beasly, right before he tells his story.]]
** [[FunnyBackgroundEvent There's something hilarious about Layton's slack-jawed expression in the background during the whole thing]], especially since when Luke looks at him he closes his mouth; but when the camera pans back to Luke and Layton, he's back to looking slightly puzzled.
* The cutscene at the top of the tower where [[spoiler:Don Paolo reveals that he has been disguising himself as Layton ever since everyone entered the tower.]] In the background, you can see Flora silently backing off without taking her eyes from him - a moment ago, she had been clinging to his arm. [[spoiler:The funny is made a little bittersweet, however, when you remember that this is the first time she's seen him since the previous game - you know, when he kidnapped her and ''took her place'' with Layton and Luke.]]
** Future Luke says that the situation is getting too complicated. [[spoiler: Which is ''hysterical'' considering the sheer complexity of the scheme he's been working on for the last few years.]]
* A simultaneous SugarWiki/MomentOfAwesome and {{SugarWiki/Funny|Moments}}, [[spoiler:when the [[CoolCar Laytonmobile]] starts flying, we get this:]]
--> '''Luke:''' Wow! We're [[spoiler:actually flying!]] Can you believe it?
--> '''Layton:''' Yes, [[spoiler:Paul [Don Paolo] was always very good with machines.]]
--> *Beat*
--> '''Luke:''' Um, Professor? [[spoiler:Just when did you learn how to fly a plane?]]
--> '''Layton:''' [[spoiler:A plane? This is an automobile!]]
* A rather funny CallBack occurs in one of the credits scenes. Looks like Rosetta got her private lesson with the Professor, all right... except not really, since of course he has Luke and Flora along with him. The look on her face just screams, "This is ''not'' what I had in mind."