The Everglades Stormwater Treatment Areas (STAs) retain phosphorus (P) from stormwater runoff before it enters the Everglades Protection Area. These large treatment wetlands are extremely efficient in reducing total P (TP) concentrations. However, in some cases, the STAs may require additional measures to reach and/or maintain mandated discharge TP concentrations. One such option, a Periphyton-based Stormwater Treatment Area (PSTA) Cell, was constructed in 2005 on 40 ha at the outflow region of one flow-way in STA-3/4. This PSTA Cell was built to determine operational and engineering requirements associated with the implementation of this technology on a large scale. The PSTA Cell has supported an oligotrophic, periphyton-rich treatment wetland over its operational period of record. This system has consistently produced annual flow-weighted mean (FWM) outflow TP concentrations at or near the mandated outflow TP levels. The PSTA Cell maintained optimal treatment performance when subjected to hydraulic conditions observed in other full-scale STA cells (i.e., deeper water depths, pulsed flows, and major storm events). The PSTA Cell's treatment performance appeared to be better (FWM outflow concentrations ∼9 μg L−1) during continuous flows for 21 days compared to intermittent flow periods. Daily PSTA Cell outflows often were ≤ 13 μg L−1 when inflow TP concentrations were ≤ 22 μg L−1. The PSTA Cell also removed additional P from very low inflow concentrations (< 15 μg L−1). Seepage into the PSTA Cell occurred when its stage was lower than stages in adjacent water bodies, but seepage was greatly reduced when stages in the PSTA Cell were increased and the difference in stage between the PSTA Cell and adjacent bodies was reduced. TP concentrations in seepage water were usually higher than the PSTA Cell's inflow or surface water TP concentrations, therefore, the PSTA Cell performance did not benefit from seepage inflows. Over the 13 water years of operation, accretion of new soil (TP < 300 mg kg−1) did not result in nutrient gradients within the PSTA Cell that might affect P performance. Overall, the PSTA Cell study achieved its goal of providing operational and engineering insights for the implementation of this technology on a larger scale, and the PSTA Cell consistently attained ultra-low-P concentrations (mean ± standard deviation) of 10 ± 3 μg L−1 over its operational period of record.