The display case currently showing the canoe features a backdrop mural of historically accurate Timucuan life painted by the artist Stacy Breheny.

[2] The main gallery consists of a permanent display of local and Florida history along the east wall, with the remainder of the space devoted to rotating temporary exhibitions.

Sarah Matheson, who died in 1996, was one of the original board members and founders of the museum and was also an early president of the Alachua County Historical Society.

The Matheson House contains original furniture, artwork and artifacts from antebellum and Victorian era Florida.

The display on the first floor consists of tools that originally belonged to the John Mason Tison Jr. or were donated by the community.

From 1976 to 1980 retired cardiologist and Matheson co-founder Dr. Mark Barrow and his wife, Mary, operated an antique store in the building.

The Matheson History Museum received a $300,000 historic preservation grant from the Florida Department of State, Division of Historical Resources, along with contributions from private donors, to transform the building into a library and archives for the museum's local history collections.