In 1994, researchers from the Florida Fish and Wildlife Conservation Commission (FWC) completed a report, entitledClosing the Gaps in Florida’s Wildlife Habitat Conservation System, assessing the security of rare and imperiled species on existing conservation lands in Florida. The biologists that authored this report used species occurrence data, habitat data, and the analytical capabilities of Geographic Information Systems (GIS) to assess the protection afforded to 62 focal species on lands managed for conservation and to identify important habitat areas in Florida that have no conservation protection. These areas, known as Strategic Habitat Conservation Areas (SHCA), depict areas needed for protection and serve as a foundation for conservation planning in Florida. Since 1994, landscape-level habitat changes, transfer of land from private to public ownership, and changes in land use have reduced the appropriateness of using the findings from the 1994 report to accurately assess Florida’s current biodiversity and wildlife conservation status. Advances in technological capabilities, revised habitat data, and more extensive species-occurrence data allowed us to reassess Florida’s biodiversity protection status. Additionally, advances in population-viability modeling techniques allowed us to examine the security of species given their current distribution, habitat needs, and the amount and distribution of habitats currently protected. We identified SHCA for a new selection of focal species, including many species that were in the original report. This project will help determine how habitat-protection needs have changed since 1994 and where protection efforts should be focused to ensure the long-term conservation of Florida’s wildlife.