The hosts already have one foot in the semi-final after cruising to a 3-0 victory at the Johan Cruyff Arena last week.
Match preview
Chelsea are on the verge of reaching the Champions League semi-finals for a second straight season after producing a dominant display in Amsterdam.
Lauren James opened the scoring in the 19th minute, before Sjoeke Nusken netted a brace to give the Blues a sizeable three-goal advantage in the tie.
Emma Hayes's side enjoyed another successful outing in Sunday away's game against West Ham United, with Aggie-Beever Jones and Erin Cuthbert grabbing a goal apiece to secure a 2-0 victory, which keeps them top of the Women's Super League table on goal difference.
After winning each of their last six matches, the Blues will take significant momentum into Wednesday's second leg, where they will attempt to maintain their 100% success rate in Champions League quarter-final ties.
Chelsea will certainly believe they have the defensive resilience required to prevent Ajax from mounting a comeback, having kept clean sheets in five of their previous six outings.
As a result of Chelsea's strong defence, Ajax will have to produce something special if they are to avoid disappointment in their first-ever appearance in a European quarter-final.
The Dutch side had enjoyed a memorable campaign prior to the first leg, taking 10 points from six matches to finish above Bayern Munich and Roma in a tricky group.
However, after experiencing disappointment last week, Ajax know they will have to become the first team since Man City in March 2023 to score three goals in an away game against Chelsea if they are to have any hope of progressing to the semi-finals.
Unfortunately for the visitors, they have struggled to perform on their European travels this season, after drawing one and losing two of their three away group games.
Suzanne Bakker's side will at least travel to England off the back of a positive result at the weekend, with Romee Leuchter scoring a hat-trick to lead her team to a 3-2 victory over PEC Zwolle.
Chelsea Women Women's Champions League form:
W
D
W
W
W
W
Chelsea Women form (all competitions):
Ajax Women Women's Champions League form:
L
D
W
L
W
L
Ajax Women form (all competitions):
Team News
Chelsea are still having to cope without the services of Millie Bright, Aniek Nouwen, Maren Mjelde, Mia Fishel and Sam Kerr.
Mayra Ramirez could be brought into the side after she returned from injury to feature as a substitute in Sunday's win over West Ham.
Guro Reiten, Eve Perisset and Cuthbert all dropped down to the bench at the weekend, but the trio are likely to be recalled for Wednesday's second leg.
As for the visitors, they are unable to call upon 16-year-old midfielder Lily Yohannes, who will miss the game through suspension due to an accumulation of yellow cards.
Sherida Spitse will be available for the midweek trip to the English capital after serving a one-match ban in the first leg.
Leuchter may represent Ajax's best hope of mounting a comeback, having scored 10 times in the last seven competitive matches.
Chelsea Women possible starting lineup:
Hampton; Perisset, Carter, Buchanan, Charles; Cuthbert, Leupolz; James, Nusken, Reiten; Ramirez
Ajax Women possible starting lineup:
Van Eijk; Keijzer, Spitse, De Sanders, De Klonia; N Noordam, Van Gool, Sabajo; Grant, Jansen, Leuchter