The chilling image was snapped by ghost hunters at the Grade I listed museum during an investigation - and participants also heard footsteps, eerie noises and staff at Cusworth have reported seeing strange figures in the building since the picture was taken.
The weekend ghost hunt by a team from Fright Nights East and South Yorkshire is sure to spark debate over what may have caused the ghostly handprint on a black board at the hall, which dates from 1740.
And it is not the first time paranormal behaviour has been reported at Cusworth, which is reputed to be one of Doncaster's most haunted buildings.
"Behind us was a door that led to a corridor.
"Part way down the corridor is a room that is set up as a school room with old wooden desks.
"While getting used to our surroundings and letting our eyes adjust Martin (one of our mediums) advised us of a very strong and annoyed energy.
"The energy was not annoyed with us but with his situation.
"All of a sudden there was a very loud scratching noise on the door. This went from top to bottom and everyone in the group heard it.
"This was the opposite door to where we had entered and we knew nobody was the other side.
"The group went to stand in the corridor where the noise had come from and we had various interactions.
"Most of the noises were on the doors and from in the school room.
"A desk was forcefully banged and footsteps could be heard all around. Nobody entered this room!
"After the vigil ended all guests were shocked to enter the room and see this handprint!
"As always we are sceptical so proceeded to test how our hand would look if we tried. Two things were noticeable straight away, the hand was huge, bigger than any guest's hand and our efforts had much more definition. Guests also confirmed that this handprint was not there earlier in the night and noises continued for the rest of the evening."
Cusworth Hall later shared the photo on its Twitter feed adding: "This ghostly hand print appeared on the black board in the basement during this weekends Fright Nights ESY Tour. We've also had staff reporting strange figures sighted within the hall this morning. What do you think?"