The Viola lead 2-0 from the teams' first meeting in Cremona, but they now return to Stadio Artemio Franchi on the back of successive defeats - the first of which finally ended their long unbeaten run.
Match preview
Ahead of the first leg earlier this month, Fiorentina had turned around a season bedevilled by inconsistency to win nine of their previous 10 matches and were fresh from victory over Inter at San Siro.
They therefore arrived full of confidence at Stadio Giovanni Zini, where they dictated terms to their hosts for much of the game and established a two-goal lead in the tie thanks to Arthur Cabral's first-half finish and a Nicolas Gonzalez penalty.
As a result, their fine form in cup competitions continued, as not only are the Viola contesting the Coppa Italia semis but a chance to reach the Europa Conference League final awaits in May.
Securing a 6-4 aggregate win over quarter-final opponents Lech Poznan saw Vincenzo Italiano's side overcome a collapse in the home leg to eventually advance, but their 3-2 defeat to the Polish club ended a 14-match unbeaten streak - worse was to follow last weekend.
Fiorentina's recent turnaround has carried them into the top half of the Serie A standings, but despite going two goals up within 13 minutes at Monza, they proceeded to fritter away their advantage and were ultimately beaten 3-2 by their mid-table rivals.
Such a setback may cause concern among the Franchi faithful, as though the Tuscan club remain hot favourites to meet either Juventus or Inter in the Coppa Italia final, conceding three goals in both of their last two games is a trend which cannot continue on Thursday.
The Viola's last major silverware came with Coppa success some 22 years ago, and after being eliminated by Juve in last year's semi-finals, they are intent on lifting the trophy for a seventh time.
While Fiorentina progressed past Sampdoria and Torino to reach this stage of the competition, Cremonese took a far more hazardous route to the final four, seeing off southern giants Napoli and Roma.
However, their cup exploits have not been matched by results in the Italian top flight. After more than 20 years away, the Lombardy side returned to Serie A last summer, but following Sunday's 3-0 defeat to Udinese they are eight points adrift of safety with just seven games left to play.
Despite following their first-leg defeat with consecutive wins in the league - against relegation rivals Empoli and Sampdoria - a quick return to the second tier still beckons for Davide Ballardini and co, but claiming the club's first-ever Coppa Italia would more than compensate.
Overturning their deficit this week will be a mountainous task, though, as all three of their encounters with Fiorentina this season have gone in the Viola's favour - including Cremo's first-day defeat at Stadio Franchi.
The Grigiorossi now return there on Thursday, knowing that it is a true case of win or bust - and even a one-goal victory on the night will not be enough.
Fiorentina Coppa Italia form:
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Fiorentina form (all competitions):
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Cremonese Coppa Italia form:
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Cremonese form (all competitions):
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Team News
While Giacomo Bonaventura and Josip Brekalo could remain unavailable, joining injured reserve goalkeeper Salvatore Sirigu on the sidelines, Vincenzo Italiano should have an otherwise full Fiorentina squad to choose from against Cremonese.
Lorenzo Venuti aims to return following illness, but Dodo is more likely to start at right-back in any case, with Lucas Martinez Quarta and Igor vying to partner Nikola Milenkovic at the heart of the hosts' back four.
Having opened the scoring in Cremona earlier this month, Arthur Cabral should again get the nod to spearhead the Viola's front three; Luka Jovic is the Brazilian's most probable replacement should Italiano opt for a reshuffle.
After Emanuel Aiwu was sent off in the first leg for blocking Cabral's shot with his hand, Cremonese will be without the Austrian defender on Thursday. Fellow centre-back Vlad Chiriches is also absent again.
Leonardo Sernicola, David Okereke and Felix Afena-Gyan have all scored twice in this season's Coppa Italia so far, but only the former is expected to start in Florence, featuring on the visitors' right flank.
Fiorentina possible starting lineup:
Terracciano; Dodo, Milenkovic, Igor, Biraghi; Amrabat, Mandragora; Ikone, Barak, Saponara; Cabral
Cremonese possible starting lineup:
Carnesecchi; Vasquez, Bianchetti, Lochoshvili; Sernicola, Benassi, Castagnetti, Valeri; Buonaiuto; Dessers, Tsadjout