- Randall's apparent death and especially its aftermath. Foreshadowed and then openly mentioned throughout the game up to that point, it still hits like a truck. Layton is utterly broken afterwards, so much that he needs to leave his village to overcome the grief for failing to save his friend. Granted, it later turns out that there was one more mystery Layton and Randall hadn't uncovered, but Layton's despair and grief are heart-rending.
- The BGM Override for the next puzzle (which also happens to be Puzzle #100) certainly helps keep the mood going. It really gives you the feeling that your imminent triumph over the ruins is all too bittersweet.
- And what's on he other side of the door? A mountain of gold and jewels. No grand discovery for the progression of archeology. Layton is seen touching one of the coins, and collapsing in grief.Layton: No technology. No magic, Randall. Just this. Gold. Gems. Metal and rocks. Was this worth your life?
- The ending of the "Snow Bunny" play- the bunny saves the boy he's friends with, at the cost of his own life.
Tearjerker / Professor Layton and the Miracle Mask