The influence of modified plasma membrane fatty acid composition on cellular strontium accumulation in Saccharomyces cerevisiae was investigated. Growth of S. cerevisiae in the presence of 1 mM linoleate (18:2) (which results in 18:2 incorporation to ~70% of total cellular and plasma membrane fatty acids, with no effect on growth rate) yielded cells that accumulated Sr²⁺ intracellularly at approximately twice the rate of S. cerevisiae grown without a fatty acid supplement. This effect was evident over a wide range of external Sr²⁺ concentrations (25 µM to 5 mM) and increased with the extent of cellular 18:2 incorporation. Stimulation of Sr²⁺ accumulation was not evident following enrichment of S. cerevisiae with either palmitoleate (16:1), linolenate (18:3) (n-3 and n-6 isomers), or eicosadienoate (20:2) (n-6 and n-9 isomers). Competition experiments revealed that Ca²⁺- and Mg²⁺-induced inhibition of Sr²⁺ accumulation did not differ between unsupplemented and 18:2-supplemented cells. Treatment with trifluoperazine (TFP) (which can act as a calmodulin antagonist and Ca²⁺-ATPase inhibitor), at a low concentration that precluded nonspecific K⁺ efflux, increased intracellular Sr²⁺ accumulation by approximately 3.6- and 1.4-fold in unsupplemented and 18:2-supplemented cells, respectively. Thus, TFP abolished the enhanced Sr²⁺ accumulation ability of 18:2-supplemented cells. Moreover, the rate of Sr²⁺ release from Sr²⁺-loaded fatty acid-unsupplemented cells was found to be at least twice as great as that from Sr²⁺-loaded 18:2-enriched cells. The influence of enrichment with other fatty acids on Sr²⁺ efflux was variable. The results reveal an enhanced Sr²⁺ accumulation ability of S. cerevisiae following 18:2-enrichment, which is attributed to diminished Sr²⁺ efflux activity in these cells.