Hauptcharaktere: Professor Hershel Layton - Luke Triton - Flora Reinhold - Don Paolo - Inspektor Chelmey - Barton - Dr. Andrew Schrader - Katia Anderson - Anthony Herzen - Frederick Beluga - Sammy Thunder
Sophia - Sir Anderson - Croy - Makroni - Chester - Babette - Oma Enygma - Irina - Geoff - Cnoblock - Jacques - Grousley - Hank - Ernest - Trudy - Lili - Sally - Marjorie - Conrad - Karla - Wandersen - Dorothea - Melck - Oscar - Nick - Gabe - Balsa - Wurtzer - Lopez - Myrtel - Porto - Lulu - Albert - Madeline - Remy - Mucky - Kosta - Dogey - Joseph - Rory - Lila - Junior - Felix - Niles - Duke - Hopper - Olson - Derby - Dawson - Joanie - Krantz - Stanley - Marina - Ophelia - Ray - Gregorio - Narice - Gertie - Schnurrbert - Polo - Herzog Herzen - Winston - Rätselfreunde - Mysteriöses Mädchen - Don Flora
Tiere: Hamster - Tom - Prinzessin - Glamour - Muhlene