Although perfluorooctanesulfonic acid (PFOS) has been voluntarily phased out, it remains the most abundant and frequently detected PFAS compound in biota worldwide. A deeper understanding of how PFOS enters the aquatic food web at the energetic base is needed to better characterize and predict the general patterns of PFAS trophic transfer. Research on bioaccumulation by primary producers remains limited. Because diatoms (Bacillariophyta) are often dominant constituents of aquatic biofilms, we exposed freshwater benthic diatoms (Mayamaea atomus) to a range of PFOS concentrations (0.01–100 μg/L) for 7 days in a controlled laboratory experiment to investigate PFAS bioaccumulation patterns. We quantified PFOS in water and algal matrices using liquid chromatography-tandem mass spectrometry and calculated bioconcentration factors (BCFs). Log PFOS concentrations in diatoms increased linearly with log₁₀ exposure concentration, corresponding to a sublinear relationship in arithmetic space. Consequently, BCF values decreased, from 4,831 to 174 L/kg, with increasing PFOS exposure, indicating concentration-dependent bioaccumulation consistent with higher-order organisms (e.g., Crustacea, Mollusca, Chordata). This pattern complicates the use of BCF for predictive purposes and may lead to mis-estimations of risk. Even as PFOS declines in the environment, algae will likely continue to accumulate and transfer PFOS and other PFAS to higher trophic levels.