Chamberlain, who umpired both Grand Finals in 2010 and the 2011 International Rules Series, made his senior debut in 2004 and has umpired 175 games, including 13 finals.
Umpiring director Jeff Gieschen said Chamberlain was initially scheduled for a rest this weekend after experiencing soreness in his knee, but scans revealed a more serious issue and he will now be sidelined until the 2014 season.
"Ray has done an outstanding job to return to top form this season after missing the whole of the 2012 year with a serious knee injury," Gieschen said.
"While he’s disappointed that he’s suffered another long-term injury, he is committed to making a return for next year.
"He is one of our best and most experienced umpires on the senior list and the AFL umpiring department and team look forward to having him return to full fitness again."