The Foxes have fought off Stoke City and Brentford to reach the last-16 stage and will be confident as they welcome the Seagulls to the King Power Stadium where they have already beaten them once before this season.
Graham Potter's men meanwhile beat Newport County and Blackpool to reach the Fifth Round and arrive in the game in good form having gone six games unbeaten in all competitions.
Leicester boss Brendan Rodgers has already confirmed that he intends to rotate his side, but with a realistic chance of silverware on the line, the Northern Irishman could go strong in order to securing the East Midlanders passage to the quarter-finals.
Goalkeeper Danny Ward was between the sticks in the Fourth Round win at Brentford and is likely to deputise in goal once more. Across the defence, Ricardo Pereira could be given the chance to rest as he still manages his full return from injury.
With Timothy Castagne out injured, it could provide the opportunity for youngster Vontae Daley-Campbell to make his senior debut for the side after being named on the bench in Leicester's last two games.
Options are limited in the centre of defence after it was confirmed that Wesley Fofana would be ruled out for a month, meaning that Caglar Soyuncu and Jonny Evans are the most likely candidates however Daniel Amartey is more than capable of coming into the side if needed. James Justin is also expected to make way for his promising understudy in Luke Thomas.
Moving into midfield and two more youngsters could be in line for first-team action. Khanya Leshabela is another who has been an unused substitute and could be called into action whilst promising talent Sidnei Tavares - who has been linked with the likes of Barcelona - was not in the Under-23's squad on Monday, implying he may be used on Wednesday. Both are likely to be on the bench however, with the ever-reliable Youri Tielemans and James Maddison to start as usual.
Rodgers also stated that they will look to give Wilfred Ndidi some minutes as he manages his recovery from a torn hamstring. Having played Hamza Choudhury in the last two fixtures, Nampalys Mendy seems the more likely to start this one.
The wide positions will likely see Harvey Barnes take his usual slot on the left-hand side whilst on the opposite, Cengiz Under could be given a chance to show he is up to the challenge of fighting for a regular starting spot.
Finally, despite Rodgers admitting Jamie Vardy would feature on Wednesday after his return in Sunday's draw against Wolverhampton Wanderers, it is highly unlikely the 34-year-old will start. Kelechi Iheanacho has impressed in his two most recent outings and could be in line to start a third successive match.
Predicted Leicester Starting XI: Ward; Daley-Campbell, Amartey, Soyuncu, Thomas; Mendy, Tielemans, Maddison; Barnes, Iheanacho, Under.
Brighton & Hove Albion
Brighton are looking to reach the last eight of the FA Cup for the second time in three seasons and with good form on the road of late, confidence will be high.
Graham Potter traditionally likes to set his side up in a 3-4-2-1 formation and starting in goal is expected to be back-up keeper Christian Walton who has featured in the previous two rounds for the Seagulls.
Adam Webster limped off during Brighton's midweek draw against Burnley so could be unavailable in which case Dan Burn would be likely to move to centre-back alongside captain Lewis Dunk and Ben White.
Yves Bissouma scored a stunning effort in the last round and will be important in the midfield battle at the King Power Stadium and alongside him to help in the centre of the pitch could be the experienced head of Adam Lallana as well as the hero at Anfield, Steven Alzate.
In the wing-back roles, Joel Veltman is a certain to start on the right with usual choice Tariq Lamptey out with injury whilst 19-year-old Polish recruit from Legia Warsaw Michal Karbownik could make his debut.
Into the attack and the front-three of Leandro Trossard, Alexis Mac Allister and Neil Maupay are all lively players with the latter thriving off the technical ability from Trossard and Mac Allister who are capable of creating something out of nothing.
Predicted Brighton Starting XI: Walton; White, Dunk, Webster; Veltman, Bissouma, Lallana, Burn; Trossard, Mac Allister; Maupay.
If the last meeting between the two is anything to go by, this should be a routine win for Leicester but the Foxes have struggled to gain momentum at home this season whilst Brighton have picked up some impressive results on the road in recent weeks.
Despite the form of both, in the cup that goes all out the window and is down to quite simply who wants it more on the day. Leicester have strength in depth and with a chance to win a trophy, Rodgers won't want that to slip away whilst for Brighton, their focus may be more focused on securing Premier League survival for this year.
Prediction: 2-0 Leicester.