Italian international Mattia Cassani has failed to recover in time for Fiorentina's trip to Juventus on Tuesday night.
The right-back was hoping to be fit enough to feature after missing the 2-2 draw with Catania because of a muscle injury.
There were whispers circulating that the on-loan Palermo man would return, but he's not been named in the 20-man squad.
Alberto Gilardino is in the squad again after he returned to action at the weekend following a spell on the sidelines.
Fiorentina squad: Boruc, Neto, Pazzagli; De Silvestri, Gamberini, Nastasic, Natali, Pasqual; Behrami, Cerci, Kharja, Ljajic, Lazzari, Munari, Romizi, Romulo, Vargas; Gilardino, Jovetic, Silva.
Think you know your Italian football? Share your knowledge, tips and comments to win cash prizes in OLBG's tipster competition  - £5,000 monthly.