The competition's been revamped and the Spireites take their first step in the Checkatrade Trophy as they welcome Wolves' under 23 side to the Proact on Tuesday.
The game is the first of three group stage games for Chesterfield, with a home match with Accrington Stanley and a trip to Crewe Alexandra coming up to see which two teams will make it out of the group.
For further details of the competition lay-out, click here
The rules of the competition mean that Danny Wilson must field five of the starters from Saturday's match with Millwall in his starting line-up again on Tuesday night.
This means that while certain members of the first-team squad are expected to be rested, there will still be plenty of regulars involved again against Wolves.
The Championship opponents on the other hand will be fielding a squad of under 23 players, though they may also include a number of first-team players in their line-up.
Among the players expected to make the trip to Derbyshire are goalkeeper Jon Flatt, who had a non-playing loan spell with the Spireites in the spring of 2015 as back-up to Tommy Lee.
After their 3-1 defeat at home to Millwall on Saturday, the Spireites will be looking to put their two recent losses to one side and start their Checkatrade Trophy campaign on a winning note.
The Wolves under 23s are managed by former Blackburn and Bolton Wanderers midfielder Scott Sellars, who was previously head of academy coaching at Manchester City.
Sellars's side have won two and lost one of their three games so far this season, their latest outing a 3-1 home win at Molineux over rivals West Brom, where Bright Enobakhare scored twice.
From the Dugout
Ahead of the clash, manager Wilson talked about the expectations he has for his side on Tuesday evening and throughout the rest of the tournament.
"There will be changes but we won't be expecting to lose the game. We'll put a team out that we expect to win the game, we want to win every game that we play in.
"There's an expectation anyway in every game that we play, be it a cup game or a league game, we're at home and we expect to be in the top areas of this mini league that we're in.
"Our approach will be no different from any other game, the preparation will be very similar."
Wilson gave details on the Wolves side he anticipates coming up against, saying that however young the side is the West Midlanders will be no pushovers.
"We know one or two of the boys that might play, and there are one or two that we don't know," he commented. "You're going to get energy, you're going to get legs and enthusiasm.
"You will see at times that they might not have a bit of know-how and game knowledge and that's hopefully where we'll come into our own.
"It won't be easy. Under 23s sounds like young ones, but there are a lot of players that are playing in the first team at under 23 anyway."
Whether you're at the game or not, you can follow the Proact action with live commentary on PlayerHD, as well as with comprehensive coverage across the Chesterfield FC official media sources.