After a disappointingly early exit from Europe's premier club competition in 2011/12, there will be a sense of unfinished business among the players when they take to the field in a sold out Signal Iduna Park.
Dortmund have been in fine fettle of late and Saturday's dominant 3-0 victory over Bayer 04 Leverkusen streched their unbeaten run in the league to an astonishing 31 matches.
Robert Lewandowski opened his Bundesliga account for the season, while the form of German sensation Mario Götze has been nothing short of exceptional after his return from injury. German player of the season Marco Reus will also be anxious to impress in what will be his first taste of top level European club fooball.
Group of champions
Despite their domestic dominance, Jürgen Klopp's troops are aware that international competition requires a different style of play. Nonetheless, BVB are full of confidence, with defender Neven Subotic commenting that Dortmund "know exactly what we're capable of." They will need to harness that ability to progress from what looks to be the toughest group of them all. Each of the four sides - Real Madrid and Manchester City make up the numbers - won their national league last season.
Ajax have been impressive themselves, scoring 16 goals in their five league games so far and have former Bundesliga star Ryan Babel back in their ranks after he returned to the club from 1899 Hoffenheim. The Amsterdam outfit also boast an enviable European pedigree, having and lifted the European Cup on three occasions.
What may play on the Dortmund players' minds are the unpleasant memories of their 2011/12 campaign. Despite their superb form in the Bundesliga, Klopp's side were unable to produce on the international front and were eliminated from the competition with just four points from their six group games, even missing out on third place and a spot in the UEFA Europa League.
While Dortmund are certainly eyeing the knockout stages, their first aim will be to show that they have learned their lesson from last season. Tuesday evening will provide an interesting test of their progress.
Dortmund: Weidenfeller - Piszczek, Subotic, Hummels, Schmelzer - Gündogan, Kehl - Blaszczykowski, Götze, Großkreutz - Lewandowski
Ajax: Vermeer - van Rhijn, Alderweireld, Moisander, Blind - Schöne, Poulsen, de Jong, Eriksen - Sana, Boerrigter