England goalkeeper Jack Butland fractured his ankle in Saturday's victory over Germany.
The Football Association announced the extent of the injury on Sunday evening, confirming in the process that the 23-year-old has left the England squad.
It means Butland is likely to be ruled out of England's plans for Euro 2016, with the tournament just 10 weeks away.
The FA said in a statement: "Goalkeeper Jack Butland will miss Tuesday night's fixture against the Netherlands at Wembley Stadium after fracturing his ankle.
"The Stoke City man sustained the injury in the first half of Saturday's 3-2 victory over Germany in Berlin and has now returned home."
The FA said England manager Roy Hodgson would not be adding to his squad at present.
England had a light training session at their Grove hotel base in Watford on Sunday as they began preparations for the Holland game, following the impressive 3-2 win in Berlin.
Butland had earlier taken to Twitter to give an update on his situation, before learning he had suffered the fracture.
He wrote: "Firstly, what a night to be involved in! an incredible and much deserved win against the World Cup winners, makes me proud to be English!
"But my nature to not give up was my downfall, the injury I'll recover from but more importantly the experience I'll learn from!
We'd like to wish @JackButland_One a swift recovery from his injury.
- England (@england) March 27, 2016
"thank you for all your messages, I'll be having a scan today to see the damage to my ankle and will work hard to get back fit! thank you!"
With Joe Hart injured, Butland was handed his fourth cap for the clash with Germany, and was replaced by Fraser Forster moments before half-time after going down in agony.
Burnley's Tom Heaton is also in Hodgson's current squad and could have a role to play against Holland.
Butland sustained the injury just before Toni Kroos opened the scoring in Berlin in the 43rd minute.
He had the chance to put the ball into touch to receive treatment, but chose instead to punt down field, whereby Germany claimed possession and raced forward through Real Madrid midfielder Kroos, who beat Butland with a low 25-yard shot at his near post.
England conceded a second to Mario Gomez in the second half but shocked the world champions as Harry Kane and Jamie Vardy struck to level before Eric Dier headed home a last-minute winner.
Manager Hodgson hailed the quality of England's comeback, and the attacking flair rather than the blood and guts often associated with famous Three Lions performances of the past.
Nights such as the gritty 0-0 draw in Italy which saw Glenn Hoddle's side qualify for the 1998 World Cup or the 1-0 win over Argentina at the 2002 World Cup in which England played with such battling passion live fondly in the memory -- but this comeback in Berlin's Olympiastadion was different.
"All credit to the players to come back and win the game in the way they did," Hodgson said.
"It was by playing football and there was no element of just rolling our sleeves up and winning more challenges -- we had been winning challenges right the way through the game.
"It was about using the quality we think this team has to create more chances and I think three goals, we have got to say with that number of chances, is a fairly meagre return."
Hodgson's team on Saturday night were certainly not shackled by previous failings of the national team -- just 161 caps were shared among the starting XI before kick-off.
But the innocence of youth, with 19-year-old Dele Alli again starring alongside the likes of Kane and Dier, both 22, was something which Hodgson never feared would lead to a loss against the vastly more experienced world champions.
"We honestly believed that, although the team was young and inexperienced, if we could carry out our game plan we could cause Germany problems and that showed from the very first minute," he told BBC Radio 5 Live.
"At half-time we had nothing to show for it . The second goal came at a time when we were playing well and looking to get back into the game and that was a cold shower for us.
"I think the thing that is going to please me the most is the general performance of the team from the first minute to the last.
"It is very satisfying that the performance then gives you a result because they don't always, and sometimes we have been on the end of games where we haven't been bitterly disappointed by the performance but we have been by the result."
If many saw this as vindication for Hodgson's continued faith in young players -- with the likes of John Stones, Raheem Sterling and Ross Barkley hoping to push their way into his side before the start of the European Championship - the 68-year-old himself does not quite agree.
"I don't know what I had to vindicate as such," he said.
"I've not felt, with the approach we've had from changing the team from 2012, 2014 to this 2016 edition, that people weren't behind that or that we were going down the wrong track or making mistakes.
"They may have raised a few eyebrows, putting Dele Alli in the team when he's not even played for the Under-20s or Under-21s and only a few games for the Under-19s.
"But we had good experiences before with people like Raheem Sterling, who we put in at an early age, or Danny Welbeck. We tried it with them and that didn't let us down.
"Not at any stage of the game did we sit back and accept that Germany would press us back, we were pressing them back more and it is so nice when you start to take your goal chances.
"There was a moment I feared I would be talking and saying the performance was good and we created chances but we didn't take them and as a manager that is always the frustrating thing because that is icing on the cake which makes a performance what it is."