The two teams have been drawn into Group F alongside Inter Milan and Slavia Prague, and it is shaping up to be a very interesting battle to secure the top two positions in the section.
Dortmund will head into Tuesday's contest full of confidence following an impressive 4-0 win over Bayer Leverkusen in the Bundesliga on Saturday. Marco Reus scored twice for the hosts, while Paco Alcacer and Raphael Guerreiro were also on the scoresheet during a fine performance.
BVB suffered a 3-1 loss to Union Berlin in their last game before the international break but returned to winning ways in impressive fashion to occupy second spot in the table - one point off leaders RB Leipzig.
Now the German club will switch their attention to the Champions League, and it will be fascinating to see how they perform in what is a tough group considering Inter's summer business and new-found belief under serial winner Antonio Conte.
This is actually Dortmund's 14th Champions League campaign - they won the competition in 1997, and claimed a runners-up spot in 2013. What's more, they have reached the round of 16 in five of their last six participations, meaning that it would be something of a surprise if they did not advance from this section.
Last season, the German outfit were eliminated by Tottenham Hotspur in the round of 16, which included a 1-0 loss on home soil. Surprisingly, Dortmund have only won three of their last 10 European home matches, which is not the best record to bring into a clash with one of the finest sides in Europe.
That said, the Bundesliga giants have only lost three of their 15 home matches against Spanish opposition, winning five of the last seven. Last season, BVB recorded an impressive 4-0 home success over Atletico in the group stages, although they did lose the away clash 2-0 in the Spanish capital.
Dortmund have plenty of recent experience when it comes to facing Spanish opposition, but they have actually never locked horns with Barca in the Champions League. Indeed, their only two meetings came in the UEFA Super Cup back in 1998, where the La Liga outfit ran out 3-1 winners on aggregate.
Lucien Favre's side will be feeling good about themselves following an impressive victory over Leverkusen, and a win over Barca on Tuesday night would suggest that the team are capable of being a force in the competition this season.
Recent form: WWWLW
Like Dortmund, Barca will enter the match off the back of a strong league win having recorded a 5-2 home success over Valencia on Saturday night. The Spanish champions were once again without Lionel Messi but managed to put three important points on the board.
Barca have already dropped points in two La Liga fixtures this season, having lost to Athletic Bilbao and drawn with Osasuna, but Saturday's win has seen them rise into fourth spot in the table. It was also another special night for 16-year-old Anssumane Fati, who scored his second goal for the club.
Luis Suarez also came off the bench to score twice on his return to the team, and the fact that the Uruguay international is now fit and available is a huge boost to the Catalan giants ahead of this match.
Messi is yet to play this season due to a calf problem, and Barca boss Ernesto Valverde recently admitted that the Argentina international is unlikely to be available for the clash with BVB. Suarez's return is huge, but the Spanish champions are just not the same team when their talisman is not available.
Barca, who won their fourth La Liga title in five years last season, have not failed to qualify for the first knockout round of the Champions League since the 2000-01 campaign. They reached the semi-finals of last season's competition after overcoming Lyon and Manchester United in the round off 16 and quarter-finals.
Valverde's side looked destined to make the final after a 3-0 first-leg win over Liverpool, but the Premier League club incredibly recorded a 4-0 success at Anfield to advance. Barca have actually only won four of their last 13 away matches in European competition, which will hand Dortmund confidence.
The La Liga side, who are five-time European champions, have won only two of their last seven matches against Bundesliga opposition on the road. Losing to Dortmund on Tuesday night would not be the end of the world for Barca, but as mentioned above, they are in a tricky group with Inter certainly showing signs of confidence under new head coach Conte.
Inter actually travel to Camp Nou on matchday two in the Champions League, and there would be huge pressure on Barca to pick up a positive result if they did suffer a defeat in Germany next week.
Recent form: LWDW
Alcacer, who has scored 10 goals in eight matches for club and country this season, will lead the Dortmund line against the club that he represented between 2016 and 2019.
Axel Witsel came through 78 minutes against Leverkusen after recovering from an adductor injury, and the experienced midfielder is expected to get the nod on Tuesday night.
Manuel Akanji also came through the Leverkusen clash despite suffering an ankle injury on international duty, meaning that the centre-back should make the XI.
Jadon Sancho has also had a fine start to the season for both BVB and England, and the in-demand 19-year-old is expected to start in the three behind Alcacer, which will also include Reus.
As for Barca, Messi is unlikely to be available. The Argentine could well travel to Germany with his teammates, although a spot in the starting XI seems almost impossible having not yet played a minute this season.
Suarez is expected to start having made his return off the bench against Valencia, though, and there will also be a spot in the final third for Antoine Griezmann.
Carles Perez has developed into a key player for Valverde early in the season, and the attacker could be given the nod over Fati, despite the 16-year-old enjoying a brilliant start to the campaign.
Valverde also has decisions to make in midfield with Sergi Roberto, Arthur, Ivan Rakitic and Arturo Vidal battling for the position alongside Frenkie de Jong and Sergio Busquets in the middle of the park.
There are not expected to be any changes further back, though, with Nelson Semedo keeping his spot at right-back after coming through 90 minutes against Valencia despite a slight ankle complaint.
Borussia Dortmund possible starting lineup:
Burki; Hakimi, Akanji, Hummels, Guerreiro; Delaney, Witsel; Brandt, Reus, Sancho; Alcacer
Barcelona possible starting lineup:
Ter Stegen; Semedo, Pique, Lenglet, Alba; De Jong, Busquets, Roberto; Griezmann, L Suarez, Perez
Head To Head
As mentioned, these two teams have only locked horns twice in European competition, with both of those matches coming in the UEFA Super Cup back in 1998. Barca recorded a 2-0 home win before claiming a 1-1 draw in Germany to record a 3-1 aggregate success.
We say: Borussia Dortmund 2-2 Barcelona
Without Messi, Barca might well be underdogs heading into Tuesday night's clash. Suarez's return is extremely important, though, and we fancy the Spanish giants to claim a welcome point at Signal Iduna Park.