The contest started on Margaret Court Arena at 12:23am, the latest ever a match has begun in Melbourne and ended at 3:12am with the Spaniard emerging victorious in the second latest finish in tournament history.
Muguruza takes the first set behind one break of serve
The match began with what would be a rare occurrence in this one: a break of serve as Muguruza got to 15-40 with a searing forehand winner and was on top quickly when Konta sent a backhand long.
The Brit was quickly under fire in her next service game, seeing a 40-0 lead pegged back to deuce as the Spaniard was hammering the ball early, but Konta managed to hold to stay in touch at 2-1.
After a pair of love holds, it was the world number 38 who applied the pressure this time, earning her first break point of the match only to see the 18th seed wipe away the chance with an ace as she would go on to hold.
Konta was hitting the ball almost as solidly as her more established opponent and that helped her fight off another threat from Muguruza as she held to continue to trail by just one break at 4-3.
The level from both was continuing to be at a high standard and it was the Spaniard who earned a set point on the Brit's serve, but Konta gamely fought off Muguruza's advances with a forehand that just clips the line.
Serving for the set, the 18th seed fell behind 15-30 only to win the next two points to bring up set point and she thumped a serve to force Konta to knock a backhand return long to claim the early lead.
Konta hits back to win second set tiebreak
Konta was the one in the ascendancy early in the second set, holding to love and getting to deuce straightaway on the Muguruza serve, but the Spaniard again got herself out of trouble with her serve.
The Brit was upping her level and it seemed to be having an effect on Muguruza, who pushes a forehand long to give Konta a break point. She screws a forehand wide and the Spaniard again escapes, this time with a backhand lob.
The 18th seed seemed to be making progress on the Konta serve as she was again at 30-30, but the former semifinalist does enough to keep her nose in front at 4-3.
Both ladies were playing and hitting the ball at an exceptional level and Muguruza was threatening to end the night early as she led 15-40 on Konta's serve only to commit a backhand error on her first break chance and the Brit drilling a backhand winner on the second to eventually hold.
The set would eventually be decided in a tiebreak which was briefly stopped after the first point to close the roof as rain began to fall.
Once the action resumed, Konta claimed the crucial minibreak with a precise backhand to go ahead 4-2, eventually winning the set on forehand that just stayed in, this instant classic heading to a third set.
Spaniard claims victory at 3:12am in classic
The final set began at 2:25am as Muguruza opened up the decider with a hold, firing an ace on game point. Konta was matching the Spaniard shot for shot as she responded with an easy hold of her own.
The Brit then reached deuce on the 18th seed's serve, but was unable to make any further progress. Same in the next game as she was at 30-30 only to see an ace from Muguruza and a backhand of her own fired wide.
Konta was in trouble on serve for the first time in a while trailing 3-2, falling behind 15-30 in her service game only to rescue the situation with a confident smash and backhand on game to point to get to 3-3.
This incredible tilt continued on at even as both ladies held to 30 and again in their next service games holding even easier to finely balance the set at 5-5. At this point, it was anyone's guess as to how this would turn out.
Muguruza held to 15 for a 6-5 lead and was two points from victory at 15-30 only to see Konta scoop an amazing forehand winner down the line to get back to 30-30. The Spaniard responded in kind on the next two points.
A fizzing return brought up match point and a final blow, a backhand winner by Muguruza sealed her place in the third round against Timea Bacsinszky and brought one of the great matches in tournament history to a close.