He fractured a vertebra in his spine when kneed in the back by Juan Zuniga in a challenge that went unpunished.
Neymar, 22, was in tears when he was carried off on a stretcher before being taken to hospital in Fortaleza.
Coach Luiz Felipe Scolari said: "Neymar was going to be hunted. For three matches that has been happening."
Team doctor Rodrigo Lasmar told Brazil's SportTV: "Unfortunately, he's not going to be able to play. He is very, very sad.
"It's not serious in the sense that it doesn't need surgery, but he'll need to immobilise it to recover."
Lasmar added that he expected the recovery period to take at least a few weeks.
Neymar recovered from thigh and knee injuries sustained in the second-round win over Chile to play against Colombia.
The Barcelona forward has featured in all five of Brazil's matches at the 2014 Fifa World Cup and is their top scorer with four goals.
Neymar at the World Cup
Matches played5
Minutes played457
Attempts on target72.2%
Goals scored4
Assists1
Neymar had a quiet game on Friday and was replaced in the 88th minute after Napoli defender Zuniga challenged him for a header, with referee Carlos Velasco Carballo taking no action.
Scolari said: "Not even a yellow card, nothing.
"People think Germany, these or the others, only they are hunted, but not Neymar."
Zuniga insisted: "I never meant to hurt a player. It was a normal move.
"I was playing for the shirt from my country, not with the intent to injure. I was just defending my shirt."
Thiago Silva's early goal and a stunning David Luiz free-kick in the second half set up victory for Brazil, who withstood a late Colombia onslaught that saw James Rodriguez score an 80th-minute penalty.
Brazil committed 31 fouls during a scrappy game while their South American rivals made 23.
Hosts Brazil meet Germany in the semi-final in Belo Horizonte at 21:00 BST on Tuesday.
Brazil captain Silva will also miss that game through suspension after picking up a second booking of the tournament against Colombia.
The final will be held at the Maracana in Rio de Janeiro on Sunday, 13 July.
Brazil v Germany will be live on BBC One on Tuesday, 8 July from 20:30 BST
Neymar was challenged by Napoli defender Juan Zuniga late in the game
The forward was in obvious pain after the incident
Brazil will be without top scorer Neymar and suspended captain Thiago Silva against Germany
Brazil fans gathered outside the hospital where Neymar is being treated
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