The grainy clip, uploaded to video sharing website YouTube, shows lights which "started to slowly move and disappeared" above the Town Hall, according to the web user who recorded the footage.
The video, entitled "lights above town hall sheffield!" was uploaded onto the web earlier this week by a user called "naisian1."
It is not clear exactly where and when the footage was shot - although a flickering white light can be seen dancing around in the centre of the screen during the early part of the video.
User "naisian1" wrote in the comments section: "Stayed there for about 3 mins, I stopped filming thinking they were possibly some cranes Newley (sic) erected In the distance or something... Then a helicopter flew towards them, they started to slowly move and disappeared?"
The 53 second clip shows vehicles moving through the city centre in darkness. There is no commentary on the footage.
Earlier this month, mysterious lights were spotted in the skies over Sheffield, sparking suggestions of aliens, Batman and supernatural forces.
Social media was awash with sightings of the searchlights which were spotted in the skies above various areas of the city in early October.
The beam spotlights, reaching high into the night sky, were spotted from Dronfield to Manor and Eckington to Mosborough.
One spotter, Laura Roberts wrote: "Can anyone else see lights over Manor way in sky? Like torch light on sky? Second night on trot ya can see it n wondering what it is."
Meanwhile, Twitter user Coops wrote: "Anyone else see these weird lights in sky in Sheffield?!"
There have been suggestions that the lights could be an alien invasion, the signal used to call Batman, ghostly goings on or even the Northern Lights.
But other, more likely explanations are lights used to promote a local travelling circus while others have indicated the searchlight beams may be being projected from a city centre night club.