Barca will make the trip to Madrid following the international break on October 14 without fans, the La Liga leaders announced on Wednesday.
Applications for the match have been cancelled as Atletico cannot supply the tickets previously offered at the newly-built stadium, though 62 members who already made a purchase are guaranteed to attend.
It comes after Barca defeated Las Palmas 3-0 behind closed doors at Camp Nou last week amid political tensions over the Catalan independence referendum.
The iconic stadium was empty after the Barca board decided to play the fixture without fans after their request to have the match postponed was rejected by LaLiga.
Barca's appeal for postponement came amid violent scenes during Catalonia's independence referendum on Sunday, which was declared illegal by Spain's constitutional court.