Whichever side comes out on top at De Grolsch Veste will progress to the playoffs, leaving them just one step away from the competition's new-look group phase.
Match preview
A late goal from Michel Vlap gives Twente real hope of turning around their third-round tie, as the midfielder's 90th-minute free kick in Salzburg reduced their first-leg deficit to only one goal.
Having trailed to a Maurits Kjaergaard brace heading towards stoppage time last Tuesday, coming away with just a 2-1 defeat sets the stage for a potential comeback on home turf.
Joseph Oosting's side finished third in the Eredivisie last season, to gain entry into the Champions League qualifiers, and they kicked off their latest league campaign at the weekend.
Summer signing Sam Lammers scored the opening goal in a 2-1 win at NEC Nijmegen, as Twente bounced back from defeat in Austria four days earlier.
The Tukkers now hope to take one step closer to a first Champions League appearance in 14 years, but even if they find a way past Salzburg, a playoff against either Rangers or Dynamo Kyiv would then stand in their way.
Regulars in Europe's top club competition during the past decade, Salzburg were eliminated at the group stage last year, finishing fourth behind Real Sociedad, Benfica and Inter Milan.
Under new management this season, Die Roten Bullen believe they have something to prove, as after a decade of dominance they finally failed to win their domestic title in 2023-24.
Losing out to Sturm Graz on the final day cost former boss Gerhard Struber his job, and ex-Liverpool assistant Pep Lijnders has returned to front-line management following a brief spell in charge of NEC several years ago.
His start to life in the top job has been successful, as Salzburg completed pre-season unbeaten, then progressed in the Austrian Cup, before a 3-2 win at Grazer AK secured three points from their first Austrian Bundesliga fixture.
Having since toppled Twente and then cruised to a 5-1 league victory over Blau-Weiss Linz, it seems normal service has been resumed ahead of this week's second leg.
FC Twente Champions League form:
L
FC Twente form (all competitions):
Red Bull Salzburg Champions League form:
W
Red Bull Salzburg form (all competitions):
Team News
While Joseph Oosting opted to rest Ricky van Wolfswinkel at the weekend, instead fielding Sam Lammers up front, Twente's captain should now return to an attack that averaged over two goals per game in last season's Eredivisie.
Sem Steijn finished as the Tukkers' top scorer last term, and he is set to start in support of Van Wolfswinkel on Tuesday evening.
Though Mees Hilgers returns from a domestic suspension and should start in defence, Czech midfielder Michal Sadilek is still sidelined by injury.
As is their model, Salzburg have said goodbye to several departing stars this summer, following the departures of Luka Sucic (to Real Sociedad) and AC Milan-bound Strahinja Pavlovic.
Furthermore, Ivory Coast international Karim Konate - who scored 20 league goals last season - recently suffered a knee injury and may be out of action until September; he joins Fernando, Takumu Kawamura and Alexander Schlager on the treatment table.
In Konate's absence, Petar Ratkov should lead the visitors' attack, supported by another hot prospect, attacking midfielder Oscar Gloukh.
FC Twente possible starting lineup:
Unnerstall; Reeger, Hilgers, Van Hoorenbeeck, Kuipers; Kjolo, Eiting; Rots, Steijn, Van Bergen; Van Wolfswinkel
Red Bull Salzburg possible starting lineup:
Blaswich; Dedic, Baidoo, Blank, Terzic; Bidstrup, Gourna-Douath, Kjaergaard; Gloukh, Ratov, Nene