Hampshire has been professionally active in the Canadian film and television industry since 1996. She has starred in the Canadian series This Space For Rent, Carl², and Northern Town. She played Margaret in The Life Before This, a feature film directed by Jerry Ciccoritti, which premiered at the Toronto International Film Festival in 1999.

In 2006, starred alongside Kevin Zegers and Samaire Armstrong in the 2006 romantic comedy It's a Boy/Girl Thing, in which she played the character Chanel. In 2009, she appeared in The Trotsky as Leon Trotsky's love interest Alexandra. In 2010, she starred in the Canadian indie film, Good Neighbours. Hampshire has also done voice-acting, voicing the character Misery on the animated series Ruby Gloom, Diana Barry in Anne of Green Gables: The Animated Series, Chloe Crashman in Carl Squared, Starr in 6Teen, and Alyson Malitski in Braceface.

Hampshire was born c. 1982 in Montreal, Quebec.

On 23 May 2012, in Cannes, France, Birks presented the first Birks Canadian Diamond award to Hampshire and fellow Actress Sarah Gadon during Telefilm Canada's inaugural Tribute To Canadian Talent press event and reception. Also in 2012, Hampshire starred in David Cronenberg's Cosmopolis as Jane Melman, and co-starred in Sean Garrity's comedy film My Awkward Sexual Adventure, for which she received positive critical notice. The same year she was cast in the futuristic-zombie film The Returned.

In 2014, Hampshire was cast in the Syfy series 12 Monkeys in the recurring role of Jennifer Goines, a reimagined version of Brad Pitt's character from the film on which the series is based; in 2015, she was upped to a series regular for the show's second season. Since 2015, she has played Stevie Budd in the CBC television series Schitt's Creek.

In late 2015, Hampshire was cast in Xavier Dolan's independent film The Death and Life of John F. Donovan, which also stars Jessica Chastain, Kathy Bates, and Susan Sarandon.