The classic track - which was a number one hit in both 1975 and 1991 - beat Michael Jackson's 'Billie Jean' and Adele's 'Someone Like You' in the ITV1 poll.
The Nation's Favourite Number One aired on ITV1 last night (July 15), after viewers had voted online to mark the 60th anniversary of the charts.
However, voters were not able to choose any number one from the past 60 years, but rather a pre-selected shortlist of songs, which included Dizzee Rascal's 'Bonkers', and just one song from the 1950s.
Oasis hit 'Don't Look Back in Anger' and 'Hey Jude' by The Beatles completed the top five.
John Lennon, Britney Spears, ABBA, Whitney Houston and Kylie Minogue also featured in the top ten number one singles.
A similar poll was run in 2002 by the Official Charts Company and Radio 2, with voters allowed to choose any number one from the previous 50 years.
Queen were joined by Adam Lambert for their new run of gigs together in the UK last week.
Watch the video for Queen's 'Bohemian Rhapsody' below: