It is housed in the former United States pavilion constructed for Expo 67 located within the grounds of Parc Jean-Drapeau on Saint Helen's Island.

[6][7] The two uppermost platforms held "Destination: Moon", an exhibit on NASA's space programs; spacecraft were hung from the dome's steel frame, including the Freedom 7, Gemini 7, and Apollo AS-202 capsules.

[8][9][10] On the mezzanine level, the "American Spirit" exhibit displayed a wide variety of artifacts, including American Indian crafts, folk art objects, guitars owned by famous musicians, a collection of dolls, and an array of almost 300 hats, representing a range of regions and occupations.

[20][21] The pavilion reverted to its nature theme in 1972, with the addition of a troop of baboons, a Japanese garden, and a children's adventure area, Sleeping Beauty's Fantasy Land.

[26] After the fire, the city was determined to maintain the Biosphere and continue using it as an open-air structure, possibly containing suspended gardens or a concert venue.

[30][31] In August 1990, Environment Canada committed $17.5 million to turn Biosphere into an interactive museum showcasing and exploring the water ecosystems of the Great Lakes-Saint Lawrence River regions.

It offers interactive activities and presents exhibitions about the major environmental issues related to water, climate change, air, ecotechnologies, and sustainable development.

[35] In 2021, control of the Biosphere was transferred from Environment Canada to Space for Life, the City of Montreal's complex of nature museums.

The sun-shading system was an attempt by the architect to reflect the same biological processes that the human body relies on to maintain its internal temperature.

[44] Disney designer John Hench expanded on Biosphere's three-quarter-sphere to create a full sphere appearing to be balanced on legs.

[45][46] The Biosphere appears in the game Civilization VI (in the New Frontiers DLC) as a World Wonder, where it increases the appeal of marsh and rainforest tiles, and boosts renewable power and tourism.