With Joe Mauer and Trevor Plouffe on base and no outs in the 12th, Colabello came through with a single up the middle to score both of them. Mauer had started the rally with an infield single before Plouffe doubled down the left-field line.
It was another big hit for Colabello, who also crushed a two-run homer as part of a four-run Twins fourth inning. He has 26 RBIs on the season to rank second in the Majors to Miami's Giancarlo Stanton (27).
"It was a big night for Chris Colabello," Twins manager Ron Gardenhire said. "He's up to 26 RBIs this month. He still has a chance to add on, which is pretty special, especially for a guy who says, as he put it, in 'beer leagues' a couple years ago. So it's fun to watch."
Colabello also tied the club record for RBIs in April, joining Hall of Famer Kirby Puckett, who had 26 in 1994. Colabello, who played in the independent Can-Am League for seven years before joining the Twins organization in 2012, has six more games this month.
"It's quite an honor," Colabello said. "You start getting mixed in with names like that and you realize how special this game is and how special this opportunity to be here is. It's certainly something I'll remember for a long time."
It was a special performance for Colabello, as both his parents were in town from Milford, Mass., to see him play. His mother, Silvana, was celebrating her birthday, and was being interviewed by Fox Sports North when her son delivered a two-run homer in the fourth.
"That's a pretty cool moment," Colabello said. "I'm obviously blessed to have my parents in my life, and to have them along with me is pretty neat. It's weird because April 23 has always been a special day for me, hitting-wise. It seems like every time I play on her birthday something crazy happens."
Colabello's go-ahead single in the 12th made reliever Casey Fien a winner, as the right-hander tossed 2 1/3 scoreless innings, including recording the final out in the ninth with the bases loaded. Closer Glen Perkins tossed a scoreless 12th to get his fourth save.
"We did some good things offensively, but they just didn't translate into enough runs," Rays manager Joe Maddon said.
Right-hander Mike Pelfrey struggled again, as he now has yet to pitch more than 5 1/3 innings through his first four starts. He gave up four runs (three earned) on six hits and three walks over five-plus innings on Wednesday night.
"I thought it was better, but it's obviously nowhere close to where I want to be," Pelfrey said. "The best thing about tonight's game is we won."
Pelfrey was hurt by bad luck in the first inning, as Desmond Jennings reached on a broken-bat infield single with one out that hit the second-base bag. Jennings then stole second base with catcher Josmil Pinto's throw sailing into center field to allow Jennings to reach third and score on an RBI groundout from Matt Joyce.
Pelfrey didn't help his cause in the third, however, as he threw a wild pitch with two runners on and one out, allowing Ryan Hanigan to reach third. It proved costly, as Joyce brought home Hanigan with a sacrifice fly to center field to give the Rays an early 2-0 lead.
But the Twins struck in the fourth against right-hander Jake Odorizzi. Plouffe started the rally with a leadoff walk before Colabello delivered his two-run blast to tie the game at 2. It was Colabello's third homer of the year and his second in as many days.
The Twins kept the rally going, as they loaded the bases with nobody out against Odorizzi before Eduardo Escobar gave Minnesota the lead with a sacrifice fly that knocked Odorizzi from the game after just 3 1/3 innings. Reliever Brandon Gomes came in and surrendered an RBI single to Sam Fuld, who was playing his first game with the Twins since being picked up off waivers from the A's on Sunday.
But the Rays promptly answered in the fifth on a two-out RBI single from Evan Longoria, who beat out a soft chopper hit to Plouffe at third base. Tampa Bay tied it in the sixth, when Pelfrey gave up a leadoff single to Wil Myers and hit Logan Forsythe with a pitch.
Michael Tonkin replaced Pelfrey, and he issued a one-out walk to load the bases, which led to the Twins bringing in left-hander Caleb Thielbar to face Ben Zobrist, who tied the game with a sacrifice fly to center. But the Rays were held scoreless over the final six innings in a game that lasted nearly five hours.
"Guys came in and made some big pitches to get out of it," Gardenhire said. "Casey was fantastic. And most of the guys who went in there did a good job. The game could've went a lot of ways, but fortunately we came through with some big hits at the end to get a big win."