Valencia are two-time runners-up in the competitionBBC coverage
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Gian Piero Gasperini's Atalanta have no injury worries as they prepare for their first ever Champions League game in the knockout round on Wednesday, in their debut season in the competition.
"This is a game that we have been waiting months for," said the Italian.
"The Champions League is a dream which we'll try to carry forward as much as possible."
Valencia will be without suspended defender Gabriel Paulista for the last-16 first-leg tie at Milan's San Siro.
The fixture, like Atalanta's other home games in Europe this season, will take place at the San Siro - home of AC Milan and Internazionale - because their Gewiss Stadium in Bergamo, which is 55km away, does not meet Uefa requirements.
Gasperini added: "Being able to play in front of so many of our fans at a venue like San Siro is a beautiful achievement that we have been waiting a long time for.
"The whole city of Bergamo is practically travelling to Milan."
His side surprised many by finishing third in the Italian top flight last season, and they come into this match sitting fourth in Serie A, having defeated closest challengers Roma on Saturday.
The match at the San Siro will be officiated by an all-English team, with Michael Oliver selected as the referee.
MATCH FACTS
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This will be the first encounter between Atalanta and Valencia. It's also the first time the team from Bergamo will face Spanish opposition in a competitive fixture.
Valencia have won only one of their eight away games against Italian opposition in the Champions League (D2, L5), a 1-0 victory against Roma in the 2002-03 second group stages. They have also failed to find the net in six of those eight games.
Atalanta are the first Champions League debutants to make it to the knockout stages since Leicester City in 2016-17.
Under the current format (since 2003-04), Atalanta are the first team to qualify for the last 16 after losing their opening three group games.
Atalanta won two of their six games in this season's group phase (against Dinamo Zagreb and Shakhtar Donetsk), the lowest tally alongside Lyon among the 16 remaining teams.
Valencia are in the knockout stages of the Champions League for the first time since 2012-13. It's been 13 years since they last made it past the last 16 (2006-07).
Valencia have never won an away game in the Champions League knockout stages (D4, L5). They've also only ever kept one clean sheet in those nine outings, a goalless draw against Leeds United at Elland Road in the semi-finals back in 2000-01.
78% of Valencia's goals in the group stages came after the 60th minute (7/9), the highest proportion among the 16 remaining teams.
Source: bbc.com