Pliskova was down 5-1 against Williams in the final set before the seventh seed produced a fightback for the ages to claim a 6-4 4-6 7-5 quarter-final win over the American great.
The Czech could muster only 20, half of which came in the second set when she hung in to level the match despite the barrage of winners from her opponent. In that game, Pliskova failed to take any of her five game points.
Osaka has now won 59 straight matches when winning the first set and also trumped Pliskova on the service front.
"In this kind of stage of the tournament, when you just go deep, from Brisbane, lot of things happen".
Reflecting on the encounter two years later, the Spaniard told the ATP: "It was a really emotional match. I'm sure he's not feeling great about ending the Grand Slam this way", said the Serb.
For Williams, all roads now lead to Roland Garros where she will be hoping for a more favourable draw.
"First foot fault of the match... on match point". How can an ankle roll occur in next rally?
Kvitova, who reached a career-high No.2 in 2011, must go one round better than Osaka for a chance of rising to No.1.
Fan Dan Leach tweeted: 'Can't believe that Serena lost that match. She went for her serves.
A foot fault is when a player over-steps the baseline while serving.
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The star did not protest and went on to lose the match. "Williams was robbed!" one fan wrote after the game alongside a video.
Williams rolled her ankle on match point and eventually surrendered her serve to allow Pliskova back into the match. "I've got balls made in the match, nothing wrong". I don't think it had anything to do with my ankle, per se.
"She just played lights out on match point, literally hitting lines", Williams said. "She literally played her best tennis ever on those shots", she added.
The 37-year-old American did not call for a trainer during the match and later would not blame the ankle for the way everything changed down the stretch, saying afterwards that it "seems to be fine". Williams chasing a record-equaling 24th Grand Slam title, that will tie her with Margaret Court, said she was looking at winning. I was nearly in the locker room, now I'm standing here as the victor. One she hit an ace, unreturnable serve. And together they've been responsible for the ousting of the likes of two-time Slam champion Garbine Muguruza, 2016 Australian Open champion Angelique Kerber, and 23-time major victor Serena Williams to get to this far in the tournament.
Surprise semi-finalist Danielle Collins' cousin flew all the way from Minnesota to be in her box for her quarter-final match against Anastasia Pavlyuchenkova. As ever, the tournament is less dramatic for her absence, especially as the defeat ruled out a rematch against Osaka.
This defeat is the earliest in Australia for Williams since 2014, when she exited with a fourth-round loss to Ana Ivanovic. This was only her eighth main WTA Tour event since she returned early a year ago and her first since the US Open in the early part of September.
"This is when you go psycho, when you have match point. just go bananas on it", she said.
While Serena and Ramos had an absolute fall-out and the 23-time Grand Slam champion kept screaming and shouting, Osaka was a vision of calmness on the other side.
The 21-year-old from Japan moved closer to a second consecutive major championship by parlaying her aggressive and powerful style into a 31-11 edge in winners against Svitolina.