The shooter, 40-year-old Scott Paul Beierle, then fatally shot himself, Tallahassee Police said.
He had shot six people and pistol whipped another after walking into the studio, in a small shopping centre.
Officers identified the two murder victims as Dr Nancy Van Vessem, 61, and Maura Binkley, 21.
Ms Van Vessem was chief medical director for Capital Health Plan, which provided affordable healthcare for the people of Tallahassee.
Capital Health Plan said in a statement that she was a “guiding, visionary force”. It added: “We all have been so blessed to have Nancy in our lives.”
Ms Binkley was a student at Florida State University. President John Thrasher said: “To lose one of our students in this tragic and violent way is just devastating.”
On Saturday, police urged anyone who saw “something unusual” around the time of the shooting on Friday night to come forward. Beierle acted alone.
Police chief Michael DeLeo said: “We're all very saddened and shocked by the events that occurred, but it's important people understand that there is no immediate threat outside of what has already occurred this evening.”
Erskin Wesson, 64, said he was eating dinner with his family at a restaurant below the yoga studio when they heard the gunshots above.
"We just heard: ‘Pow, pow, pow, pow.’” Mr Wesson said. "It sounded like a limb falling on a tin roof and rolling."
Melissa Hutchinson said she helped treat a "profusely" bleeding man who rushed into a bar after the incident. She said three people from the studio ran in.
"It was a shocking moment something happened like this," she said.
City commissioner Scott Maddox was on the scene. He said on Facebook: "In my public service career I have had to be on some bad scenes. This is the worst. Please pray."
Tallahassee mayor Andrew Gillum, who is the Democratic nominee for governor, broke off the campaign trail to return to Tallahassee. He earlier appeared at a campaign event with former President Barack Obama.
Tallahassee's crime and murder rate has been an issue in the governor's race. Mr Gillum's opponent, Republican Ron DeSantis, calling the capital Florida's most crime-ridden city, a claim that is incorrect.
Additional reporting by Associated Press.
