Ms Rousseff is accused of illegally manipulating finances to hide a growing public deficit ahead of her re-election in 2014, which she denies.
Senators voted to suspend her by 55 votes to 22 after an all-night session that lasted more than 20 hours.
Vice-President Michel Temer will now assume the presidency while Ms Rousseff's trial takes place.
The trial may last up to 180 days, which would mean Ms Rousseff would be suspended during the Olympic Games in Rio de Janeiro, which start on 5 August.
According to BBC News Ms Rousseff made a last-ditch appeal to the Supreme Court to stop proceedings, but the move was rejected.
Ms Rousseff, who was first sworn into office in January 2011 and started a second term in 2015, has called the steps to remove her a "coup".
In a speech at the end of the all-night Senate session, attorney general Jose Eduardo Cardozo said that the impeachment request did not have legal basis and that the opposition wanted to remove a democratically-elected president.
He said senators were condemning an "innocent woman" and that impeachment was a "historic injustice".
Michel Temer became interim President as soon as Ms Rousseff was suspended.
- The 75-year-old law professor of Lebanese origin was Ms Rousseff's vice-president and was a key figure in the recent upheaval
- Up until now, he's been the kingmaker, but never the king, having helped form coalitions with every president in the past two decades
- He is president of Brazil's largest party, the PMDB, which abandoned the coalition in March
- In recent months, his role has become even more influential; in a WhatsApp recording leaked in April, he outlined how Brazil needed a "government to save the country".
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