Lazio, meanwhile, require a win to be certain of competing in the knockout stages for the first time since the 1999-00 campaign, but should Zenit St Petersburg beat Club Brugge in the other match in Group F, both Dortmund and the Italians will progress regardless of their result against each other.
Dortmund were hugely impressive in their second successive 3-0 victory over Brugge last week.
Erling Braut Haaland scored a brace to make it six goals for the Norwegian striker in only four days, with England international Jadon Sancho's sensational free kick adding gloss to the convincing win, moving them five points clear of their opponents in third place with only two matches remaining.
Having lost 2-1 at home to FC Koln in the Bundesliga at the weekend, Lucien Favre may be tempted into resting players against Lazio in order to focus on Saturday's trip to Eintracht Frankfurt.
However, it may be a case of wrapping up qualification and top spot in the group by beating their forthcoming opponents before that comes into the 63-year-old's thinking, especially having lost 3-1 to the Italians in the reverse fixture.
However, veteran striker Artem Dyzuba struck back to give the Russians hope, before Immobile sealed his side's victory with a brace, taking Lazio four points clear of Brugge in Group F with only two matches remaining.
Given they are competing in the group stages for the first time since the 2007-08 season, progressing to the knockout stages would be a tremendous achievement for Simone Inzaghi and his side, even if the Rome-based club were arguably handed a favourable group.
However, the former Italy international will not allow his players to get ahead of themselves, especially with a final matchday showdown with Brugge still to come. A first defeat in this season's competition at Dortmund on Wednesday would turn that last fixture into a decider for both teams if the Belgians avoid defeat to Zenit.
Borussia Dortmund Champions League form: LWWW
Borussia Dortmund form (all competitions): WWLWWL
Lazio Champions League form: WDDW
Lazio form (all competitions): WDDWWL
The Dortmund boss may be tempted to rest Haaland, with the 20-year-old requiring protection from physical overload, but will likely look to get the job done on Wednesday before making a string of changes against Zenit next week should his side manage to do so.
Former Liverpool midfielder Lucas Leiva was rested in Saturday's dismal 3-1 defeat at home to Udinese, with the Brazilian likely to come back into the starting XI in order to protect the back three and fellow former Liverpool player Pepe Reina.
Borussia Dortmund possible starting lineup:
Burki; Meunier, Akanji, Hummels, Guerreiro; Witsel, Dahoud; Sancho, Reus, Hazard; Haaland
Lazio possible starting lineup:
Reina; Patric, Hoedt, Acerbi; Lazzari, Parolo, Lucas, Alberto, Marusic; Immobile, Correa
We say: Borussia Dortmund 3-1 Lazio
We can envisage both sides posing each other problems in offensive areas, but ultimately Dortmund possess the more dangerous attackers and the more reliable defence.
Favre's side should have enough about them to get the job done and qualify as winners of the group, leaving Lazio looking over their shoulders at matters in Brugge.