Matlacha Pass NWR, administered as a satellite refuge of J.N. "Ding" Darling NWR, is located within the Matlacha Pass Charlotte Harbor estuary, Lee County, Florida, approximately 8 miles northwest of Fort Myers. On September 26, 1908, three small islands were established as a ". . . preserve and breeding ground for native birds" by President Theodore Roosevelt through Executive Order 943.

Since then, the refuge has grown to 23 islands encompassing about 512 acres. The most recent addition of lands to the refuge was on April 10, 1991, when Public Land Order 6843 withdrew approximately 312 acres of public lands from surface entry and mining for 40 years for use by the USFWS. Designated by the State of Florida as an Aquatic Preserve, the refuge islands are part of a highly productive estuary which supports a diverse array of wildlife. The islands contain two basic vegetative zones, upland sand ridges and mangroves.

Fisherman Key, one of the largest islands of the refuge, once had fish camps and permanent residents on it. Several islands of the Matlacha Pass Refuge are used as roosting and nesting sites for a variety of wading and water birds. The beaches and shores provide loafing, feeding, and nesting areas for migratory ducks, shorebirds, gulls, and terns.

Ospreys, which are listed as a priority species in the Regional Resource Plan, can be observed nesting and feeding in Matlacha Pass area. Several endangered and threatened species benefit from the habitats described including: bald eagles, wood storks, sea turtles, and manatees.