The tie is delicately poised following a goalless draw in the first leg last month, with Lyon making it very difficult for the Spanish champions on the night.
It would be fair to say that the treble is very much on for Barca this season. Saturday's 3-1 win over Rayo Vallecano moved them seven points clear at the top of La Liga, while they will face Valencia in the Copa del Rey final at the end of May. Yet another domestic double is in the offing, but the Catalan giants will be absolutely desperate to land the European Cup this season.
Barca have not won this competition since 2015 and have lost at the quarter-final stage in each of the last three seasons, which is simply not good enough for a team of their ilk.
This is the 15th straight year that the Spanish giants have made the knockout round of the Champions League, though, and they have been victorious in 12 of their previous 14 ties. Last season, Ernesto Valverde's side beat Chelsea 4-1 at this stage, while their famous 6-5 aggregate win over Paris Saint-Germain occurred at the same stage of the 2016-17 campaign.
Between February 2 and February 19, Barca drew four of their five matches in all competitions and it would be fair to say that their performances were not fantastic. They won away at Sevilla in the league on February 23, though, before recording back-to-back victories over Real Madrid in the Copa del Rey and La Liga.
As mentioned, Valverde's team will enter this game off the back of a 3-1 success over Rayo in the league. It is going to take some effort from second-placed Atletico Madrid to prevent Barca from winning the league title, while they are the big favourites to beat Valencia and land another Copa del Rey crown.
This week will be the first time that Barca have welcomed Lyon for a European match in more than 10 years. The Spanish outfit ran out 5-2 winners at Camp Nou during the last-16 stage of the 2008-09 competition and have been victorious in each of their three home fixtures with Lyon in UEFA competition.
What's more, Barca have won eight of their 11 home games against French opposition, suffering just one defeat. Lyon have a lot of talent, but it is going to take some effort from the Ligue 1 side to triumph on Wednesday night.
First in Group B this season, Barca have lost just one of their last 18 Champions League matches and are unbeaten this season. Domestic matters are taking care of themselves at the moment and therefore the Catalan giants can fully concentrate on putting things right in the Champions League.
Recent form: WWDWDD
Recent form (all competitions): DWWWLW
Lyon gave an extremely good account of themselves in the first leg last month. Barca would have been looking for an advantage to bring back to Spain, but the French outfit picked up a solid first-leg result in front of their own supporters.
This is actually the first time in seven seasons that Lyon have reached the knockout round of the Champions League. The French outfit are facing their 10th last-16 tie in the competition and despite winning the first three, have lost five of their last six encounters at this stage.
Indeed, Lyon's last victory in the round of 16 came during the 2009-10 campaign, when they overcame Real Madrid 2-1 on aggregate. They were beaten by APOEL in the 2011-12 season, though, and have found it a real struggle to compete at this stage of Europe's leading competition in recent years.
Les Gones opened their group-stage campaign with a 2-1 victory over Manchester City, but then drew each of their next five to finish on eight points, which was enough to secure second spot in the section.
Lyon have actually only lost one of their last eight European away fixtures, while it is worth remembering that they reached the semi-finals of the Champions League in 2010 and are certainly capable of holding their own at this level.
Their form has been mixed since the first leg, though, losing against Monaco and drawing with Strasbourg in Ligue 1, but they did beat Caen 3-1 in the Coupe de France before putting five past Toulouse in the league earlier this month.
Bruno Genesio's side will need to put in a near-perfect performance if they are to book their spot in the quarter-finals at Barca's expense. There have already been some incredible results in this season's Champions League, though, and Lyon certainly have players capable of pulling off a shock result.
Recent form: DDDDDD
Recent form (all competitions): WDLWWD
Ivan Rakitic and Ousmane Dembele started Saturday's league clash with Rayo on the bench, but the pair are expected to return to the XI on Wednesday night with Philippe Coutinho and Arturo Vidal dropping to the bench.
Clement Lenglet is back from a domestic ban, meanwhile, and is expected to get the nod over Samuel Umtiti at centre-back as the latter is still working his way back to full fitness following a knee problem.
As for Lyon, captain Nabil Fekir missed the first leg through suspension, but the 25-year-old is available for this game, which is a huge boost for the visitors.
Fekir has scored 12 times and registered seven assists in all competitions this season and will line up in the final third of the field.
There is a chance that Genesio could decide to bring an extra midfielder into his team on Wednesday night, which would mean one from Moussa Dembele, Memphis Depay and Bertrand Traore missing out in the final third.
The fact that Depay did not feature in the league at the weekend makes the former Manchester United winger a certainty, though, and Traore seems the likeliest to drop out for the visitors.
There will once again be eyes on midfielder Tanguy Ndombele, meanwhile, with the 22-year-old linked with a number of Europe's biggest clubs.
Barcelona possible starting lineup:
Ter Stegen; Semedo, Pique, Lenglet, Alba; Busquets, Rakitic, Arthur; Dembele, L Suarez, Messi
Lyon possible starting lineup:
Lopes; Dubois, Marcelo, Denayer, Mendy; Ndombele, Aouar, Tousart; Fekir, Dembele, Depay
Head To Head
Wednesday night's clash will be the eighth meeting between the two teams in all competitions. Barca lead the head-to-head four wins to Lyon's zero, with the remaining three fixtures finishing level.
Lyon have visited Barca on three occasions; their last trip ended in a 5-2 defeat back in March 2009, while they have also lost 3-0 and 2-0 on the road to the Spanish giants.
We say: Barcelona 2-0 Lyon
Lyon showed their credentials in the first leg, but Barca have been excellent in recent weeks and we expect Valverde's side to make the quarter-finals.