The Viola are ultimately seeking a seventh national cup and lead their visitors in the league, but Bologna's last-16 victory over Inter Milan sparked dreams of a first Coppa win for 50 years.
Match preview
Last year's losing finalists, Fiorentina aim to go one step further in the Coppa Italia this season and claim the trophy in Rome, following their painful defeat to Inter in the 2023 decider.
Also beaten in the Europa Conference League final, the Viola's thirst for silverware has not been quenched, so coach Vincenzo Italiano has continued to put plenty of resources into both competitions this term.
In addition to improving their league form, Fiorentina - who last lifted the cup in 2001 - sealed progress to the Conference League's knockout phase and then edged out Parma in their first Coppa tie, played at Stadio Artemio Franchi last month.
The latter was far from straightforward, though, as the Tuscan side trailed by two goals heading into the final 10 minutes, relying on late strikes from M'Bala Nzola and Riccardo Sottil to force extra time against the Serie B leaders, who went on to lose their nerve in a subsequent penalty shootout.
Having since climbed into Serie A's top four thanks to a run of three wins without conceding a goal, Fiorentina were favourites to prevail against struggling Sassuolo on Saturday night, when victory would have drawn them level with third-placed AC Milan.
However, Andrea Pinamonti's early goal split the sides in Reggio Emilia, and Italiano's attention must quickly shift attention to his team's two remaining fixtures before flying to Saudi Arabia for the Supercoppa Italiana: Sunday's league game against Udinese and a midweek cup contest with a side they beat 2-1 back in November.
Although star striker Joshua Zirkzee pulled them level with a penalty after going behind on 17 minutes, Bologna's most recent trip to Stadio Franchi ultimately ended in defeat, with Fiorentina's Nico Gonzalez slotting a second-half spot-kick past Lukasz Skorupski to leave the Rossoblu empty-handed.
It was a rare setback for Thiago Motta's men, who had surged up the standings with a superb series of results that took them into Italy's top four until a post-Christmas wobble, which has left the Emilian club sitting just behind Fiorentina.
That fall-off in form comprised a shock 3-0 loss to Udinese - just their second defeat since the opening day of the season - and salvaging a last-gasp draw against Genoa at Stadio Renato Dall'Ara on Friday night: veteran full-back Lorenzo Di Silvestri struck deep into stoppage time to make it 1-1.
Nonetheless, Bologna dominated their visitors for much of the game and remain unbeaten in their last 10 home matches - winning eight - while beating both Cesena and Hellas Verona in the earlier rounds of the cup. Their away form, however, is less impressive.
The Rossoblu have won just once on the road in Serie A, and their most memorable away success this season was, by far, a last-16 fightback to knock holders Inter out of the Coppa Italia last month.
Zirkzee brilliantly set up both goals in the second half of extra time, as Bologna left San Siro stunned and set up this week's quarter-final tie in Florence, with a chance to then meet either Milan or Atalanta BC in the semis.
Fiorentina Coppa Italia form:
W
Fiorentina form (all competitions):
D
D
W
W
W
L
Bologna Coppa Italia form:
W
W
W
Bologna form (all competitions):
W
W
W
W
L
D
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Team News
Fiorentina will again be without wingers Nico Gonzalez and Riccardo Sottil on Tuesday, as both continue to recover from their respective thigh injuries, in addition to long-term absentees Dodo and Gaetano Castrovilli.
Another wide option, Christian Kouame, is now away with host nation Ivory Coast at the AFCON, and the Viola's recruitment team are attempting to bring in fresh blood via the transfer market, but centre-forward M'Bala Nzola turned down a call-up by Angola and will vie with Lucas Beltran to start up front.
Goalscoring defender Luca Ranieri and cup goalkeeper Oliver Christensen could come into the hosts' starting XI, and Italiano may rotate his remaining resources amid a packed fixture schedule.
Bologna, meanwhile, can welcome back versatile midfielder Lewis Ferguson, who served a Serie A suspension against Genoa last time out, though Jesper Karlsson, Adama Soumaoro and last-16 matchwinner Dan Ndoye all remain absent through injury.
It remains to be seen whether Lukasz Skorupski will replace Federico Ravaglia in goal, while Sam Beukema and Milan loanee Alexis Saelemaekers are strong contenders for promotion from the bench.
After picking up his fifth booking of the season, Joshua Zirkzee - the Rossoblu's San Siro hero in the previous round - will be suspended against Cagliari at the weekend; the ex-Bayern Munich man should therefore start in Florence.
Fiorentina possible starting lineup:
Christensen; Kayode, Milenkovic, Ranieri, Parisi; Duncan, Arthur; Ikone, Barak, Brekalo; Beltran
Bologna possible starting lineup:
Skorupski; De Silvestri, Beukema, Calafiori, Kristiansen; Moro, Aebischer; Orsolini, Ferguson, Saelemaekers; Zirkzee
We say: Fiorentina 0-1 Bologna
The latest Derby dell'Appennino sees two teams separated by a huge mountain range meeting with little to choose between them this season. Only the lack of a first-rate striker has limited Fiorentina's progress, but Bologna boast an attack featuring Joshua Zirkzee and can upset the goal-shy Viola.