An experiment was conducted to evaluate the effects of pasture finishing versus feedlot finishing, over time, on fatty acid metabolism in Angus crossbred steers (n = 24). Ruminal fluid, serum, and adipose tissue biopsies were obtained on d 0, 28, 84, and 140. Pasture forages and diet ingredient samples were obtained at 14-d intervals to determine nutritive value and fatty acid composition. The feedlot diet consisted of corn silage, cracked corn grain, soybean meal, and a vitamin and mineral supplement. The pasture-finished steers grazed sequentially on triticale (x Triticosecale rimpaui)/annual ryegrass (Lolium multiflorum), alfalfa (Medicago sativa)/orchardgrass (Dactylis glomerata), and a cool-season grass/legume mixture. The feedlot diet contained an average of 57% of total fatty acids as linoleic acid and 2% as linolenic acid. The pasture forages contained 9% of total fatty acids as linoleic acid and 66% as linolenic acid. Concentrations (% of total fatty acids) of linolenic acid were greater (P < 0.05) in ruminal fluid, serum, and adipose tissue of the pasture-finished steers, compared with the feedlot-finished steers. Concentrations (% of total fatty acids) of cis-9, trans-11 CLA were greater (P < 0.05) in adipose tissue of the pasture-finished steers than feedlot-finished steers. Concentrations of cis-9, trans-11 CLA in adipose tissue declined (P < 0.05) in the feedlot-finished steers from d 0 to 28 to 84. In the pasture-finished steers, concentrations of cis-9, trans-11 CLA in adipose tissue (mg/g of tissue) peaked (P < 0.05) on d 28 and remained elevated (ranged from 9.91 to 12.80 mg/g of tissue) throughout the duration of the study. In the pasture-finished steers, linolenic acid concentrations tended to peak (P = 0.07) on d 28 and remained elevated (ranged from 0.64 to 0.80% of total fatty acids) throughout the study. It appears that only a short time is needed to alter the n-3 and CLA composition of adipose tissue in cattle finished on pasture.