Until now, Michigan State had been too much of a hurdle for Florida to overcome since the national championship game in 2000.
The Spartans won that game for their second national title, then beat the Gators in a regular-season rematch last year.
But sixth-ranked Florida used a smothering fullcourt press to beat the 24th-ranked Spartans 74-70 Wednesday night.
"When we're playing Michigan State, I know people are talking about how it's a big rivalry," Florida coach Billy Donovan said. "But it's hard to be a rivalry when you don't win."
Matt Bonner's career-high 23 points led the Gators (5-1), while Brett Nelson added 15.
Florida had the experience over the Spartans (4-3), who lost seven players from the team that made a third straight Final Four appearance last season.
"This is a potential Final Four team," he said. "What I love about them so much is they remind me of our team the last three years."
In other games involving ranked teams Wednesday, it was No. 1 Duke 82, Temple 57; No. 11 Kentucky 99, VMI 57; No. 17 Marquette 73, Dayton 51; No. 20 UCLA 65, UC Riverside 50; and No. 25 Western Kentucky 78, Akron 68.
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