Dutchman Tom Dumoulin, who had his overall lead sliced down to around 30 seconds by Nairo Quintana on Tuesday after a toilet break, retained the Maglia Rosa as he finished safely in the peloton, who were happy to let the breakaways have their day.
Rolland broke away with Slovenia's Matej Mohoric and Russian Pavel Brutt at the start of the day and spent virtually the entire stage at the front, eventually being joined by a leading group of 25, none of whom were in GC contention.
Several riders tried to attack, including Rui Costa and Tejay van Garderen, but they could not make it stick.
The 30-year-old Rolland then seized his chance and finished 23 seconds ahead of Costa for his first Giro stage victory.
Victory means he is the only active French rider to have won stages on both the Tour de France and Giro.
"I'm so happy, I've waited for this moment for a long time," Rolland said at the finish. "It's never easy to manage a big (breakaway) group like today, it's like a lottery."
Australian Rory Sutherland was third.
Sunweb's Dumoulin takes a 31-second lead into Thursday's Queen stage in the Dolomites, which features five consecutive categorised climbs.
(Reporting by Martyn Herman; editing by Mark Heinrich)
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