The US, who beat Japan 1-0 after extra time to reach the semi-finals, face the Germans again, having thrashed them 4-1 in the group stage.
Emma Hayes' side are expected to face a greater challenge this time around as Germany, playing in their fifth semi-final in as many Olympic Games, are eager for revenge.
"You saw in the group stage they scored four goals against us. We want to give it back to them," Germany defender Giulia Gwinn said at the Lyon stadium.
"We are collectively strong, if we take our chances, we’re dangerous for every opponent."
Spain, who survived a scare to knock out Colombia on penalties after trailing by two goals, will also have a re-match against Brazil, who they beat 2-0 in the first round.
Twice silver medallists Brazil, in their sixth Olympic semi-finals, will be looking for another upset win at Marseille Stadium after knocking out the hosts despite the absence of their suspended captain and all-time leading scorer Marta.
The 38-year-old six-time Olympian Marta will also miss the semi-finals after she was handed a two-match ban following a red card for a dangerous challenge on Spain's Olga Carmona in their group-stage defeat.
"Everything we have gone through has taught us valuable lessons and helped us improve," Brazil defender Tamires said. "The team is getting a stronger and stronger mentally."
Spain, competing at their first Games, will be aiming to extend their trophy haul after winning last year's World Cup, as well as the UEFA Nations League, and become the first world champions to triumph at the subsequent Olympics.
"We know what we're capable of, we don't feel any pressure," said Spain goalkeeper Cata Coll after their win on penalties.
"What defines us is that we are ambitious. That's what drives us on to go as far as possible."