The epic Intolerance will be shown at the Cosy Cinema at the Concertina Club to mark 100 years since its release.
Visitors will experience the film with a live score by local musician Mat Handley.
The film is regarded as one of the most expensive and most ambitious silent movies ever made.
The story sees director David Wark Griffith interweave the segments of four stories over great gaps of space and time.
The villains of the four stories are mill owner Jenkins and his intolerant social reformers, the hypocritical Pharisees - opponents of Christ, the evil regime of the cunning Queen Catherine, and the treacherous High Priest of Babylon.
Their powerful actions set in motion disturbing consequences for a modern-day working-class couple.
Chris Chapman, who helped to set up the Cosy Cinema through the arts project Right Up Our Street, urged members of the public to come along.
He said: “This is going to be an amazing experience.
“We are really excited to be able to show this truly epic film with a live score.
“Mat Handley is a fantastic local musician who has created a whole new score for the film and will be presenting this live on the night.
“We are always looking for ways to expand on the cinema experience at the Cosy Cinema and we do feel that this will be one not to miss.”
The film will be screened tonight at 7.30pm.
Tickets cost £3 and are available through the Cosy Cinema’s Facebook page.
The Cosy Cinema was set up to bring back a cinema to Mexborough so film fans don’t have to travel to Sheffield and Doncaster.
Project leaders have previously held open air cinema showcases in the town centre during the summer months.
Hundreds of people gathered with popcorn to watch screenings of popular films including Disney favourite Frozen, Kes and Flash Gordon.