The preserve is accessible from two land locations: the Winkler Point entrance in Fort Myers and the Estero River Scrub entrance in Estero.

Numerous miles of trails offer visitors the opportunity to hike the uplands of the preserve and enjoy the variety of wildlife and native vegetation that the park protects. The trails in the preserve are rugged and seasonally flooded in some areas. Sturdy footwear, insect repellent, sunscreen and plenty of water are recommended.

Estero Bay Preserve is dog-friendly provided that your pet is on a six-foot or shorter leash and any waste is bagged and removed.