With just four wins apiece, both sides languish just outside the drop zone in their respective league tables, but all the pressure will sit on the hosts' shoulders.
Match preview
Losing 2-0 to a team that had last tasted success way back in September, Claudio Ranieri's ailing Roma side hit a new low on Sunday evening, painfully conceding twice during second-half stoppage time in Como.
Now winless in 15 games away from Stadio Olimpico - a run dating back to late April - the Giallorossi have slipped further down the Serie A standings, and they are now only two points above the relegation places.
In all, they have lost half of their 16 league games so far, which represents the capital club's worst tally at this stage of a campaign since 1973.
Ranieri inherited a team in turmoil, after the brief reign of Ivan Juric soon turned sour, and the Rome-born boss is clearly struggling to turn things around.
Having recently overseen his first wins since returning for a third spell in charge - against Lecce and Europa League opponents Braga - the 73-year-old was then undone by rookie coach Cesc Fabregas, so another slip-up in the Coppa Italia cannot be countenanced.
Furthermore, losing to one of his old clubs in the last 16 would be particularly painful - especially as Sampdoria have fallen even further following his departure.
Since Ranieri left Liguria in 2021, the one-time Scudetto winners have sunk down to Serie B, where they are currently closer to the relegation zone than the playoff places.
Saturday's 0-0 home draw with nearby Spezia followed a 5-1 thumping by league leaders Sassuolo, and Samp sit 15th after 17 rounds of action, having recently parted ways with former coach Andrea Sottil.
By contrast with some mediocre form in Italy's second tier, the Blucerchiati have enjoyed their run in this season's Coppa Italia, which was incidentally kicked off by knocking out Como.
In round two, Samp conquered city rivals Genoa in a dramatic Derby della Lanterna, with Fabio Borini's late leveller setting up a penalty shootout that ultimately saw them win 6-5 on spot-kicks.
During the Ligurian club's 1980s to '90s golden era, they claimed the Coppa no fewer than four times within a decade, and Leonardo Semplici's side are now just three wins away from reaching the final.
However, they must first upset the odds by eliminating Roma, and a meeting with AC Milan at San Siro will await the winners.
Roma form (all competitions):
L
D
L
W
W
L
Sampdoria Coppa Italia form:
W
W
Sampdoria form (all competitions):
L
L
D
D
L
D
Team News
With one eye on Sunday's must-win league game against Parma, Claudio Ranieri is set to rotate his squad in midweek, when stars such as Paulo Dybala and Manu Kone could be rested.
Already established names in the hosts' squad, young trio Matias Soule, Tommaso Baldanzi and Niccolo Pisilli may be considered for promotion from the bench.
Leandro Paredes recently returned to training and could feature in midfield, but Bryan Cristante is still struggling with an ankle sprain and Mats Hummels has been laid low by flu.
Meanwhile, Sampdoria's recent change of management means that their selection is even more unclear.
There are even three goalkeepers in contention - each of whom has played this season - but Simone Ghidotti played a starring role on Saturday and could keep his place at Stadio Olimpico.
Leonardo Semplici has suggested he could introduce some fresh faces on Wednesday, when experienced defensive trio Simone Romagnoli (Achilles tendon), Bartosz Bereszynski (muscular) and Alex Ferrari (knee) will be sidelined by injury.
Roma possible starting lineup:
Ryan; Mancini, Hermoso, Angelino; Celik, Paredes, Pisilli, Zalewski; Soule, Baldanzi; Shomurodov
Sampdoria possible starting lineup:
Ghidotti; Venuti, Meulensteen, Riccio; Depaoli, Benedetti, Ricci, Akinsanmiro, Ioannou; Borini, Tutino
We say: Roma 3-1 Sampdoria
Only victory will do for Roma, who are well behind Serie A's top seven and uncertain of progressing in the Europa League.
Written by
Jonathan O'Shea