After several weeks suffering a niggling injury in the right ankle due to synovitis, the Netherlands hit-man will be sent for an operation that, while not definitively ending his campaign, is likely to see him return in pre-season.
"Ruud van Nistelrooy will today be sent for an exploratory operation on his right ankle to further investigate the chronic synovitis he has been suffering," the club said on their website.
"This op will be performed in Amsterdam by ankle arthoscopy specialist Dr C.N. van Dijk, following consultaion with the medical staff of Real Madrid."
It had initially been thought that he would be able to feature after a period of rest, but he swelling worsened in recent days and the player was not even in the match squad for the defeat to Deportivo at the weekend.
Van Nistelrooy, 31, first felt pain during the defeat to Getafe three and a half weeks ago. He is Real's second top scorer with 12 Liga goals this season and is expected to miss around two months of football.
Los Merengues lead the Primera Division by seven points from Barcelona with ten games remaining.