Jordan Henderson and Harry Maguire are set to feature against Czech Republic in England's final Group D game on Tuesday night, while Jude Bellingham could be in line to replace Mason Mount in midfield.
Henderson and Maguire are yet to play at the Euros, having been carefully managed upon their return from injury, but manager Gareth Southgate wants to give the pair game time before next week's last-16 match.
Results on Monday night ensured England qualified for the knockout stages before the Czech Republic game - victory at Wembley on Tuesday would seal top spot for Southgate's side and therefore a June 29 meeting with the Group F runners-up in the last 16.
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Meanwhile, Mount and Ben Chilwell will miss England's final Group D game against the Czech Republic at Wembley on Tuesday night after being told to self-isolate up to and including next Monday.
The pair were identified by Public Health England as close contacts of Scotland's Billy Gilmour after his positive test following last Friday's match.
Mount and Chilwell embraced and spoke to Gilmour on the pitch following England's draw with Scotland at Wembley on Friday and also spent time with their Chelsea team-mate in the tunnel afterwards - a conversation which lasted around 25 minutes.
"Chilwell and Mount will isolate and train individually in private areas at England's training base St George's Park, with the rest of the squad returning there after tonight's fixture against Czech Republic at Wembley," an FA statement read.
"The entire England squad and staff had lateral flow tests on Monday and all were again negative, as was the case with Sunday's UEFA pre-match PCR tests."
Mount and Chilwell will return at Southgate's disposal for their round of 16 tie should England top their group, but will certainly miss out on the match if they come second.
Southgate is considering replacing Mount with Jude Bellingham against Czech Republic, while Raheem Sterling and Phil Foden could also keep their places in attack despite the team's disappointing performance against Scotland on Friday.
Analysis: Time for a back three?
Sky Sports News reporter Kaveh Solhekol...
Southgate was asked about the fact that the team looked tired on Friday, and he made the point that the starting XI were in perfect physical condition, which sounded like he wants to give them another chance.
Henderson and Maguire will definitely play, and Maguire is really pushing for a start ahead of Mings. Going into such a massive game next week, you can't have a situation where Henderson and Maguire haven't had some game time.
There won't be wholesale changes, it's not a case that they're through to the next round and he's going to do what Roberto Mancini did with Italy the other day. The performance on Friday was so disappointing, one of the worst under his tenure, and he wants to give a lot of those same players a chance to make up for it.
It could be time for Southgate to revert to a back three, which is the same system he used in all the games at the World Cup in Russia, now that he has all three of those centre-backs available again - Walker, Stones and Maguire. That would be a way of getting Grealish in the side, and ditching those two holding midfielders.
But, if it's another 4-2-3-1, I can't see how Grealish can play. Southgate was only able to pick his team once he had the clarification from Public Health England around Mount and Chilwell's availability, so it's a really unusual situation because in normal circumstances, the players would know the team the day before a game.
Mount was always going to start under Southgate, and the noises coming out of the England camp is that Bellingham - still 17 years old - is in contention to replace him. That would be a massive call, Southgate has been really impressed by him in training and his maturity around the camp, so he wouldn't have any problems starting him.
Maguire 'feeling good' ahead of return
Maguire has not played for six weeks due to an ankle injury, but the Manchester United captain is feeling positive ahead of his possible Euro 2020 bow.
"I'm feeling good," Maguire told the Lions' Den. "Obviously [after] the ankle injury, it's getting stronger, I'm getting more confident.
"The more sessions I do, the more games I play, I'm sure I'll get more confidence - and that's the thing with these type of injuries.
"I think it's just about building the confidence. It's all stable now and I'm available and ready to go. Any Euro match is really important to go out there and try and win the football match.
"The boys have put ourselves in a great position in the group - four points from two games - although obviously we want to win every football match, so we'd like to be on six.
"But it's still in a strong position and now it's another big game against the Czechs. It's one we're looking forward to and we go there and try to get three points."