The Groupama Stadium will host the first leg of the last-16 tie this week, before the two teams meet at Camp Nou in the return match on March 13.
Somewhat surprisingly, this is 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, they have actually lost five of their last six encounters at this stage.
Indeed, Lyon's last victory at this stage of the competition 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 the Champions League in recent years.
Les Gones opened this season's Champions League group-stage campaign with a 2-1 victory over Manchester City, but then drew each of their next five to finish on eight points; that total was enough to secure second spot in the section, though, and set up this fascinating battle with the Spanish champions.
Bruno Genesio's side are the underdogs, but they have only lost one of their last 10 European matches, while they have won seven and lost just one of their last 13 home European games. What's more, the Ligue 1 outfit have been beaten only once at this stage of the Champions League on home soil - a 2-0 reverse to Roma in 2007.
Lyon will be hosting Barcelona in the Champions League for the first time since February 2009. During that particular season they also locked horns in the round of 16 and while Lyon picked up a 1-1 draw on home soil, they lost 5-2 in the second leg at Camp Nou to exit the competition 6-3 on aggregate.
The fact that Lyon have never beaten Barcelona in a European match is a worry, but the French side have won six of their last seven fixtures in all competitions, including a 2-1 success over Guingamp in the league on Friday night.
Genesio's side - third in Ligue 1 - also beat Paris Saint-Germain 2-1 on home soil earlier this month, while there is absolutely no question that they have the talent to cause the Spanish champions all sorts of problems on Tuesday night.
It is a tough ask for Lyon to beat Barcelona over two legs, but a positive result in the first leg on home soil would certainly give them a chance of progressing to the quarter-finals of Europe's elite competition.
Recent form: WDDDDD
Recent form (all competitions): WWWWLW
While there is every chance that Barcelona could land yet another La Liga and Copa del Rey double this season, the Champions League is the trophy that the Catalan giants really want to lift come the end of the campaign.
Indeed, the famous club 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, Barcelona beat Chelsea 4-1 at this stage, while their famous 6-5 aggregate win over PSG occurred at the same stage of the 2016-17 campaign.
Ernesto Valverde's side are without a win in their last five away knockout games, but they triumphed at both Tottenham Hotspur and PSV Eindhoven in the group stage of this season's competition, in addition to picking up a 1-1 draw at Inter Milan. A total of 14 points saw them top Group B ahead of Tottenham.
The presence of Lionel Messi means that the Catalan giants have to be seen as one of the favourites for the trophy this season. However, it would be fair to say that they have not been in the best form in recent weeks.
Indeed, Valverde's side only scraped past Real Valladolid in La Liga on Saturday night and they had drawn their last three matches in all competitions leading into the weekend fixture.
Barcelona now have a seven-point lead at the top of La Liga, but they face a huge game away at Sevilla next weekend before travelling to Real Madrid for the second leg of their Copa del Rey semi-final on February 27. March also gets off to a bang as they visit Los Blancos once again, this time in the league.
The fact that Barcelona have only won seven of their last 19 away games in the Champions League suggests that Tuesday will be a very, very difficult 90 minutes for the Spanish champions.
Recent form: WWWDWD
Recent form (all competitions): WWDDDW
Lyon will be without their captain Nabil Fekir for the first leg, with the 25-year-old missing out through suspension. The Frenchman has 11 goals in all competitions this season and his absence is a big blow for the hosts.
Marcelo, though, should be fit after only suffering from cramp in Friday's Ligue 1 fixture.
Memphis Depay has scored six times and registered 13 assists for Lyon this season, and the former Manchester United attacker will be keen to put on another strong performance against a top-level European club.
Samuel Umtiti is also unlikely to feature as he battles back from a knee problem, but there will be a number of changes from the team that started against Valladolid at the weekend.
Luis Suarez, Ivan Rakitic, Nelson Semedo and Clement Lenglet will all return, but Philippe Coutinho could be left on the bench once again with Ousmane Dembele expected to get the nod in the front three.
Lyon possible starting lineup:
Lopes; Marcal, Marcelo, Denayer; Mendy, Aouar, Ndombele, Tete; Depay, Dembele, Cornet
Barcelona possible starting lineup:
Ter Stegen; Semedo, Pique, Lenglet, Alba; Busquets, Vidal, Rakitic; Dembele, L Suarez, Messi
Head To Head
Tuesday will bring just the seventh meeting between these two teams in European competition. Barcelona have won four of the previous six, remaining unbeaten in the process.
As mentioned, the Spanish giants recorded a 6-3 aggregate win when they met in the last-16 stage of the 2008-09 competition, but all four of their other meetings were in the group stages.
We say: Lyon 1-1 Barcelona
We are expecting a very tight match in France. Barcelona will feel that any score draw is a good result, while Lyon will be looking to stay in the tie ahead of the second leg at Camp Nou.