After weeks of speculation about his future, Puyol announced at a press conference on Tuesday afternoon that he would be bringing the curtain down on his title-littered, 19-year career at the Nou Camp at the end of this season.
The 35-year-old Spain international had a contract with Barca until 2016 but has reached an agreement with the club to rescind the final two years of the deal.
Puyol, who has struggled with injuries in recent years and has managed only 12 appearances in all competitions this season, said: "After two very serious operations, I'm finding it very hard to get back to the level I need to be at to stay here - much more than I thought and much more than the surgeons told me. That's why I have made this decision.
"I'm not going today. There are three months of the season left and anyone that knows me will know that I won't give up and I'll keep fighting to the end to try to improve and help the team to achieve its objectives this season."
Puyol has made 593 appearances for Barca since making his first-team debut in 1999 - second only to current team-mate Xavi in the club's history - and helped the club win 21 trophies.
Among them are six La Liga titles, three Champions League crowns, two Spanish Cups and two FIFA Club World Cups.
The veteran central-back was also a pillar of the Spanish national side, making 100 appearances for La Roja between 2000 and 2013.
Puyol was a member of Spain's triumphant teams at Euro 2008 and the World Cup two years later, although he missed the side's successful title defence at Euro 2012 with a knee injury.
As to what he plans to do after leaving the Nou Camp, Puyol admits he is not sure yet - save to take a break.
"I don't know what I'll do after June 30, but what I am sure of is that when the season ends I'm going to need to get some rest. I haven't stopped for four years because of injuries. After that, we'll see what happens," he said.
"I would like to have a press conference at the end of the season to say a proper goodbye after a total of 19 years at the club."
Puyol's announcement led to a flood of tributes and appreciations from the world of football.
Barca goalkeeper Victor Valdes, whose time at the Nou Camp is also drawing to an end, wrote on Twitter: "There will never be another like him. Great captain! Great@Carles5puyol!"
Their club and international colleague Andres Iniesta added: "I will continue to enjoy playing by your side for the rest of the season, an honour. Afterwards, the best for you! No-one like you capi!"
Marc Bartra, who could be the long-term replacement for Puyol at the heart of the Barca defence, tweeted: "Thanks for all that you've done for me. For your help and your advice. Three more months learning by your side!"
Barca board member Pilar Guinovart, meanwhile, said that Puyol was "an example to follow for future generations".
Many more of Puyol's team-mates also added their praise, as did some of his former colleagues, such as Celta Vigo's Andreu Fontas, who said of his fellow defender: "You are the greatest: Idol, example, eternal, reference, legend, captain, friend. Thank you Puyi."
Abelardo, who played for Barca between 1994 and 2002, said in El Mundo Deportivo: "Puyol has been for several years the best defender in the world.
"He will be remember as one of the best in history and for me will be remembered as the most important defender in Barca's history."
Former Italy stars Franco Baresi and Demetrio Albertini also added their acknowledgements with the latter believing Puyol will have no shortage of offers in the summer.
Albertini told Efe: "Puyol is a great friend and has a place in any team. If he goes, it will be to succeed elsewhere."