Looking to overturn a 4-1 deficit in the quarter-final tie, Eusebio Di Francesco's side face a truly daunting task. But, with key players back and 60,000 passionate fans ready to cheer the side on, can the team produce another miracle?
At the Camp Nou six days ago the Giallorossi had two compelling penalty claims, saw their players score three times... and yet still somehow came away having lost 4-1.
Edin Dzeko's late away goal undoubtedly lends the tie a different complexion as the two sides head back to Italy, but Di Francesco's side will perhaps do the best to put the rest of the game out of their minds. Unfortunate own goals from Daniele De Rossi and Kostas Manolas gave Ernesto Valverde's side a commanding, if perhaps undeserved, early advantage - with Gerard Pique and Luis Suarez subsequently scoring to set up the current situation.
Despite the final scoreline, Roma came away from the game with great credit - and will hope to take some of the more encouraging lessons from the 90 minutes into this all-important second leg.
Both teams have played once since their last meeting - with Roma succumbing to a 2-0 home defeat against Fiorentina on Saturday afternoon.
Later that same day, Barcelona continued their march towards the Spanish title with an emphatic 3-1 win over Leganes - as Lionel Messi grabbed a hat-trick.
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Roma boss Eusebio Di Francesco has received two big boosts ahead of the match: with Radja Nainggolan back to fitness after missing the first leg with a knock (the Belgian played 90 minutes against Fiorentina on Saturday), and Cengiz Under fit enough to at least be included in the matchday squad.
Diego Perotti is absent after picking up an injury at the Camp Nou, however, while Rick Karsdorp remains a long-term absentee.
Barcelona now have no concerns over the fitness of Messi, while Philippe Coutinho is ineligible. The big doubt surrounds Sergio Busquets, whose participation will be a matchday decision.
Andres Iniesta, Samuel Umtiti and Jordi Alba all skipped the weekend win over Leganes - suggesting they will be utilised at the Olimpico.
Eusebio Di Francesco: "I think we should choose our strongest line-up tomorrow, it’s important and we must believe. Let’s perform with great heart and passion. We have to honour and respect our club colours, so we’ll give it our all and we deserve the chance to achieve something huge.
"Why not believe until the end and hope we can achieve something unprecedented? We’re trailing by three goals, so it’s a really tough ask, but we have the duty to try. So, our best team will start tomorrow and then we’ll think about the derby.”
Ernesto Valverde: “I’m expecting Roma to approach the game with great intensity, especially at the beginning. They will want to get the first goal to give themselves a chance at qualifying. And we need to play a great game to get through the tie and look forward.
“We have to approach the game not thinking that we have already qualified, despite the good result we got at the Camp Nou. We are ahead, yes, but we have to play as if it were still 0-0. Because anything can happen in a Champions League match.”
It goes without saying that Lionel Messi is Barcelona's most dangerous and compelling threat - but this is one game where Barcelona can perhaps get by without his goals. The Spanish side will progress as long as they concede fewer than two goals, meaning defensive talisman Gerard Pique could have a decisive role to play.
Valverde may be blessed with an array of great defensive options, but Pique's blend of experience, organisational skills and leadership make him the absolute lynchpin of the side. If Busquets misses the game, as some suspect, the back four may be slightly more exposed than usual; this Pique will have a huge role to play.
Strong in the air, calm on the ball and decisive in the tackle, Pique transmits confidence to his team. But he is capable of being knocked off his game and, if Roma can manage that, it could be a key to getting back into the tie.
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How much did Roma miss Radja Nainggolan in the first leg? The answer may never be known, but the Belgian midfielder at least as a chance to offer some hints with a big second leg performance at the Olimpico on Tuesday.
The return of Radja is a huge boost for Di Francesco: adding both another goalscoring option to the side and greatly enhancing the high-pressing game that is so crucial to Roma's tactical approach.
Without Busquets in the centre Barcelona might - just might - be a bit vulnerable, and Radja could be the player to make the most of that weakness. An all-action performance from the all-action midfielder could have huge ramifications for the game.
With Radja Nainggolan back and fit, Di Francesco's remaining selection decision would appear to be in attack - where, depending on the fitness status of Cengiz Under, he has a number of options to choose from in support of Edin Dzeko.
Without Diego Perotti, Stephan El Shaarawy, Cengiz, Patrik Schick and Gerson would perhaps all seem to be in contention for the two wide spots - with Di Francesco hinting at his pre-match press conference that Schick could be an option, with the Czech forward having "goalscoring in his blood".
Without Sergio Busquets, Valverde might opt to stick Paulinho in central midfield, with Sergi Roberto and Andres Iniesta alongside Ivan Rakitic to offer a midfield both compact in defence and capable of keeping the ball for long spells.
Elsewhere, given the rotation seen at the weekend, it would seem Valverde is leaning towards selecting the same side that procured Barcelona's current advantage.
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