Defeat to Inter on Matchday 1 leaves Spurs facing an uphill struggle in Group B.
But how much do you know about these two ahead of a potential UCL thriller?
Did you know that these two have only met once before in a competitive fixture? And that it was over 30 years ago?
Regulars Barca are now in their 23rd UCL campaign, but Tottenham remain relative newcomers - this is only Spurs' fourth appearance in the competition.
And did you know that Barcelona have played three previous European Cup/Champions League ties at Wembley?
Take our tricky quiz below and put your knowledge to the test – and then share it with your mates to prove just how much you know ahead of a Champions League mega-fixture.