He has appeared on television, film, and radio in the UK in productions such as Holby City, Doctor Who, Hut 33, Peep Show, The Thick of It, Keeping Mum, Fate: The Winx Saga, and The Inbetweeners.

[citation needed] He was educated at St John's School, Leatherhead, from 1990 to 1992[2] and left to receive a first in English literature at Collingwood College at Durham University.

[3] Macqueen played sarcastic consultant anaesthetist Keith Greene in 75 episodes of Holby City between May 2005 and July 2010.

[citation needed] Macqueen acted in a series of videos on YouTube as Clive Rudloe, a character who claims to be "the world's number one DJ".

[citation needed] In 2010, he appeared as the publisher Malcolm Dodds in Woody Allen's You Will Meet a Tall Dark Stranger.

He again returned to the role in The Stuff of Legend, an audio drama which was also staged as a live theatrical performance at Cadogan Hall in September 2024.

In 2016, he acted opposite Sir Michael Caine in the film Youth, and played the role of Patrick Jarvis MP in series 3 of Peaky Blinders.