The Gators compete in Division I of the National Collegiate Athletics Association (NCAA) and are single-sport members of the Big 12 Conference, which they joined after the 2024 season.

They play their home games in Donald R. Dizney Stadium on the university's Gainesville, Florida campus, and are currently led by head coach Amanda O'Leary.

The Gators have won regular-season conference titles in 11 of the 13 completed seasons of the women's lacrosse program's existence,[a] with four each in the ALC and Big East plus three in The American.

The University Athletic Association (UAA) quickly proceeded from new idea to reality in the creation of its newest sports program—it decided to create a women's lacrosse program in 2005, publicly announced the new program in 2006, hired a head coach and joined a conference in 2007, and announced its first recruiting class and began construction of a dedicated lacrosse stadium in 2008.

In October 2017, The American announced that it would start a women's lacrosse league in the 2019 season, with Florida and Vanderbilt joining four full conference members.

[8] The first Gators recruiting class of 24 players[9][10] included seven US Lacrosse first-team high school All-Americans—Caroline Chesterman, Katie Ciaci, Brittany Dashiell, Samantha Farrell, Janine Hiller, Jamie Reeg and Julie Schindel, and US Lacrosse named another five as All-American Honorable Mentions—Ashley Bruns, Lelan Bailey, Jenna Hildebrand, Hayley Katzenberger and Mikey Meagher.

[11] Dashiell, Bruns and Farrell, plus Kitty Cullen, Colby Rhea and Haydon Judge, were also named Under Armour All-Americans.

[14] The young Gators finished their inaugural season 10–8, including ALC wins over Penn State and Johns Hopkins,[3] and were ranked eighteenth in the country in LaxPower's final power ratings.

[19] The young Gators eventually fell to the veteran Duke Blue Devils 13–9 in the quarterfinals ("Elite Eight") of the 2011 NCAA Tournament, finishing their second season with an overall win–loss record of 16–4.

[25][26] The Gators defeated the Albany Great Danes 6–4 in the opening round of the tournament, and overwhelmed the Penn State Nittany Lions 15–2 in the quarterfinals.