Rosie

Rosie (puppeteered and voiced by Rebecca Nagan) is the female rag doll. During their adventures, she carries a bag that has her name written on it. She wears a yellow dress and has long shaggy black hair. She and Jim often refer to the boat driver as "fizzgog" (John Cunliffe), "loopy lobes" (or just "loopy"; Pat Hutchins) and "tootle" (Neil Brewer). She sometimes calls Jim "Noggin". Her skin colour became a bit darker from series 2.

Jim

Jim (puppeteered and voiced by Robin Stevens) is the male rag doll. In their adventures, he can be seen carrying a notebook, in which he sometimes draws things that he has seen. He has short red hair and wears light brown trousers, a shirt with a red scarf, a white-with-red-stripes sock, brown boots and a brown waistcoat. The shade of his skin has changed slightly throughout the series.

Duck

Duck (also voiced by Robin Stevens) is an animatronic wooden model of a duck, who sits on top of the Ragdoll boat. He does not talk but can be heard quacking and flapping his wings when there are no humans around to let both Rosie and Jim know that the coast is clear. In some episodes, either Rosie or Jim give him a kiss. He is the only character to directly interact with all three presenters (as only Cunliffe directly addressed Rosie and Jim).

Narrowboat owners

Rosie and Jim's home is the narrowboat Ragdoll. Throughout its run the boat was acquired by three owners, each one playing a role according to their real-life profession.

• John "Fizzgog" Cunliffe: 1990–1991, 50 episodes

An author who writes stories about Rosie and Jim based upon his own everyday outings. Cunliffe is the only presenter to break the fourth wall, delivering pieces to the camera and narration throughout every episode, which ended with a self-penned short cutout animated story done by Alan Rogers and Peter Lang (creators of Pigeon Street). He is the only presenter to directly address Rosie and Jim on-screen.

• Pat "Loopy Lobes" Hutchins: 1995–1996, 45 episodes

An illustrator who creates pictures, gathering ideas from her adventures. Each episode would start with Pat finishing a drawing of Rosie and Jim and end with an illustration, again by Pat, based upon the episode. At the end of each episode, Duck would be hidden in the picture, with Rosie and Jim trying to find him. Pat also illustrated the books and artwork for the video releases between 1995 and 1996. Hutchins is the only female presenter of the series.

• Neil "Tootle" Brewer: 1997–2000, 80 episodes

A singer and musician who plays the harmonica and the concertina. Neil would begin each episode looking for his harmonica followed by Rosie and Jim talking about and introducing him. Brewer would often end each episode with a song played on his concertina; prompted by Rosie and Jim who would leave the instrument for him to play, next to Duck. At the end of each episode, the rag dolls would often join in with the song, unknown to Neil. Brewer became the longest-serving owner of the boat and was the only one to star in a live theatre tour of the show in 1999 and 2000. Brewer is the only presenter to have starred in more than 2 series. He is also the last surviving presenter, following the death of Pat Hutchins in November 2017, and that of John Cunliffe in September 2018.

Following the boat's redundancy from Ragdoll Productions, it is no longer on display to the public.