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Gilchrist County Sheriff Bobby Schultz identified the slain deputies during a news conference as Sgt. Noel Ramirez, 30, and Deputy Taylor Lindsey, 25.
The deputies were getting food at the Ace China restaurant in Trenton when the shooter walked up to the building and fired at them through a window, Schultz said. Fellow deputies responding to the scene found the shooter dead outside the business.
Schultz wouldn't say how the suspect died, adding that state law enforcement officials were investigating. There's no apparent motive for the shooting.
"Sgt. Ramirez and Deputy Lindsey were the best of the best," Schultz said. "They were men of integrity, men of loyalty. They were God-fearing, and they loved what they did, and we are very proud of them."
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Schultz said he rushed to the scene as soon as he heard about the shooting. He said it's important to remember that attacks like Thursday's can happen anywhere, even in small cities like Trenton.
"After 26 years of doing this, there is nothing that can prepare you for senseless deaths," Schultz said.
The shooter's name wasn't immediately released.
Trenton is in northern Florida, about 35 miles west of Gainesville, and about two hours north of Tampa.
The Holmes County Sheriff's Office, in the Florida panhandle, wrote on social media that: "about an hour ago, two deputies with the Gilchrist County Sheriff's Office were shot and killed in a suspected ambush. Please keep the families of these deputies, Sheriff Bobby Schultz, and the rest of their law enforcement family in your prayers."
The Associated Press and ABC News contributed to this report.