Pine Island Flatwoods Preserve totals nearly 920 acres and is made up of 8 separate acquisitions purchased in 2000, 2001, 2002, 2003, 2005, 2008, and 2011. The southern portion of the Preserve is bordered by Stringfellow Road to the east, mangrove swamp and mudflats to the west, a future residential development to the north, and residential homes to the south. The southern portion contains the public access and hiking trails. The most recent acquisition of the Preserve is about 1/2 mile north from the larger southern portion. The latest 190 acre acquisition (nomination 389) is on the east side of Stingfellow Road and extends through a mangrove forest out to Matlacha Pass; and is now referred to as Pine Island Flatwoods Preserve - Bayside.
The Preserve consists of nine native plant communities, including mesic pine flatwoods, freshwater marshes, tidal swamp, hydric hammock, coastal grassland, coastal berm, improved pastures, abandoned groves, and unconsolidated substrate.