Located at the confluence of Olustee Creek and the Santa Fe River, Santa Fe Spring is the most upstream Floridan Aquifer spring along the river. Depending on water levels, the spring is approximately 70 feet deep in places, and it is also extremely dynamic in levels, clarity, and flow.

Santa Fe Spring is located in the 254 acre Santa Fe Springs Preserve, an effort between Alachua Conservation Trust and SRWMD.

No public access is currently available. The spring is expected to open to the public in late 2023 with multi-use trails.