Most of the 20 million items it contained, including the oldest human remains ever found in the Americas, are believed to have been destroyed.
The cause of the blaze is not known. No injuries have been reported.
The museum, located in a building that once served as the residence for the Portuguese royal family, celebrated its 200th anniversary this year.
The fire started on Sunday evening, after the facility had closed for the day.
Aerial images broadcast on Brazilian television showed it spreading throughout the building.
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