Andrej Kramaric gave the visitors the lead against the run of play shortly before the hour mark, which would have been enough for Croatia to win the group themselves while consigning England to relegation.
Find out how all of the action unfolded courtesy of our minute-by-minute updates below.
It is all to play for today, with any of the three teams in Group A4 able to finish top going into this final match. England and Croatia ultimately have things in their own hands today, but both will be expecting very difficult tests here...
Well, there are some interesting selections from the hosts, due partially to a minor injury for Jordan Henderson which prevents him from winning his 49th cap this afternoon.
Henderson is actually the most experienced member of the squad now that Wayne Rooney has left once again, but it is still a more experienced tea today than it was against USA on Thursday.
Delph is likely to play in a slightly more advanced position this time around, with Ross Barkley completing the midfield trio, but it is interesting that the Manchester City man has been selected ahead of the likes of Alli, Loftus-Cheek and Sancho.
Rashford was battling with clubmate Jesse Lingard for a place in the starting XI tonight, but Southgate has gone for Rashford in the end.
Ben Chilwell therefore starts at left-back, with Kyle Walker coming in on the right as expected as Trent Alexander-Arnold drops out despite his goal against USA.
Nikola Vlasic comes into the side in place of Rakitic - who has returned to Barcelona - which also results in a switch of formation from 4-3-3 to 4-2-3-1.
Ante Rebic led the line in that match but is expected to play in a more reserved role today, with Kramaric expected to start as the one striker.
Tin Jedvaj does start once again, meanwhile, having scored twice in the victory over Spain - including a 93rd-minute winner.
This will be the third time that England have faced Croatia in 2018, and on paper it looks like their best chance to win one of those too.
Any of the three teams in the group could still finish top, while either England or Croatia are guaranteed to be relegated this afternoon, making this a truly must-win game.
However, a goalless draw would see Spain win the group and Croatia finish third on goal difference, whereas a score draw would see England finish bottom and suffer relegation.
La Roja looked to have things wrapped up after only two games - particularly following their 6-0 win over Croatia - but successive defeats have really opened things wide open in this group.
This time last year England played out goalless draws against Germany and Brazil which failed to excite fans at all, but now it is all to play for in this game.
Reaching the World Cup semi-finals will obviously be the headline for their year, but beyond that England have made great strides with a young team and their win over Spain in Betis was among their most memorable of recent times.
Southgate's side still managed to cruise to a comfortable victory, though, with Wilson, Dunk and McCarthy all making their debuts and Wilson and Alexander-Arnold getting their first international goals.
That is one of just two defeats England have suffered from their last 27 matches on home soil, with 21 of those ending in victory.
As is the case with England, a victory would send Croatia through whereas anything else will not do for them. A goalless draw would see Croatia relegated.
Just 12 months ago they played out a goalless draw with Greece to book their place in Russia, and since then it has more or less been improvement the whole way for Croatia.
Zlatko Dalic's side have lost to Peru, Brazil, France and Spain this year, and only narrowly beat Jordan last month to end a four-match winless streak.
A victory today would really cap off a special year for the Croats.
Croatia have won just one of their last 11 away games too, failing to score in six of those including their most recent away outing - the 6-0 defeat to Spain.
One slight concern may be how much longer this team can keep on going for, with Mandzukic now retired, Modric and Rakitic the wrong side of 30 and other approaching that age too.
This should be a very interesting game, and it looks very even on paper too. We are backing the hosts to eke out a small victory. though.
SPORTS MOLE SAYS: England 2-1 Croatia
Croatia won on that occasion - via extra time - to break England's hearts just one game short of their 1966 moment, whereas in the reverse fixture of the Nations League they played out an eerie 0-0 draw.
that is England's only previous win over Croatia at Wembley, though, with a draw in 1996 and that famous defeat in 2007 which prevented England from qualifying for Euro 2008. England's only other win against Croatia on home soil saw then win at Portman Road.
ENGLAND STARTING XI: Pickford; Walker, Gomez, Stones, Chilwell; Delph, Dier, Barkley; Sterling, Kane, Rashford
CROATIA STARTING XI: Kalinic; Vrsaljko, Lovren, Vida, Jedvaj; Modric, Brozovic; Rebic, Vlasic, Perisic; Kramaric
Kalinic can only get enough on it to take it to Kane, who sees his first effort cleared off the line by Jedvaj and the second saved by a retreating Kalinic.
.@England have started well at Wembley but they should be ahead!
Harry Kane is unmarked in the 6-yard box but his volley floats high and wide. 🎈
Watch @England v Croatia in the UEFA Nations League live on Sky Sports Football now or follow: https://t.co/mpLuCpvLs3 pic.twitter.com/Vmdnitc9Z8— Sky Sports Football (@SkyFootball) November 18, 2018
The hosts have played some really good football in this match and have created plenty of chances, but the only thing missing so far has been a goal.
Sterling has also missed a one-on-one chance, and Southgate will be urging his players to be more clinical in the final third during the second half.
Perisic also had a shooting chance five minutes before half time, but the hosts will certainly need to show more in the second half if they are to avoid relegation.
A low pass into the box finds Kramaric, who has his back to goal and initially it looks as though he has to give it to Perisic. However, he keeps hold of it, twisting and turning and leaving Stones on the deck before seeing his shot deflect off Dier before looping past Pickford.
Kane had initially taken a free kick down and poked it past the keeper, and Lingard was waiting there to complete the job.
2 - Jesse Lingard has scored as many goals in his last two games for England (2) as he had managed in his first 20 caps for the Three Lions. Hope. #ENGCRO pic.twitter.com/ft2DwfKiAK— OptaJoe (@OptaJoe) November 18, 2018
A free kick is swung in from the left and Kane once again shows his poacher's instincts by sliding in to turn the ball past the keeper.
Croatia took the lead through Kramaric, but goals from Jesse Lingard and Harry Kane saw Gareth Southgate's side turn it around in thrilling fashion.
Thank you very much for joining Sports Mole for today's match as England come from behind to beat Croatia and book their place in the Nations League semi-finals. I will leave you with our match report, and be sure to stick around for reaction too.
From me, though, it is goodbye for now!