Our objective was to determine effects of dietary high-oleate (Oleate; 76% 18:1) or high-linoleate (Linoleate; 78% 18:2) safflower seeds on fatty acids in muscle and adipose tissue of feedlot lambs. White-faced ewe lambs (n = 36) were fed a beet pulp, oat hay, and soybean meal basal diet (Control), blocked by BW, and allotted randomly to dietary treatments. Cracked safflower seeds were used in isocaloric and isonitrogenous replacement of beet pulp, oat hay, and soybean meal so that Oleate and Linoleate diets contained 5.0% additional fat. Fatty acids were determined in semitendinosus, longissimus dorsi (longissimus), and adipose tissue from the tail head (tailhead adipose tissue), adjacent to the 12th rib (s.c. adipose tissue), and kidney and pelvic fat (KPH adipose tissue) depots. Fatty acid data were analyzed within muscle and adipose tissue as a split-block design. Single degree of freedom orthogonal contrasts were used to compare treatment effects. Average daily gain, feed efficiency, and carcass characteristics did not differ (P = 0.15 to 0.96) across dietary treatments. Adipose tissue saturated fatty acids were greater (P = 0.04) for Controls but were not different (P = 0.36) in muscle. Trans-vaccenic acid (18:1(trans-11)) increased (P < 0.0001) with safflower supplementation and was greater (P < 0.0001) in Linoleate than in Oleate for both tissue types. Linoleate lamb had greater (P < 0.0001) PUFA than Oleate lamb in muscle and adipose tissue. Conjugated linoleic acids (CLA; cis-9, trans-11 and trans-10, cis-12) were greater (P < 0.0001) in muscle and adipose tissue of lambs fed safflower seeds. Lambs fed Linoleate had greater (P < 0.0001) CLA in adipose tissue and muscle than lambs fed Oleate. Saturated fatty acids were greater (P < 0.0001) in s.c. adipose tissue than in tailhead adipose tissue. Mono- and polyunsaturated fatty acids were greater (P < 0.0001) in tailhead adipose tissue than in s.c. adipose tissue. Weight percentages of 18:1(trans-11) ranked tailhead adipose tissue = KPH adipose tissue > s.c. adipose tissue and semitendinosus > longissimus, whereas CLA ranked tailhead adipose tissue > s.c. adipose tissue > KPH adipose tissue and semitendinosus > longissimus. Feeding mono- and polyunsaturated fatty acids increased tissue 18:1(trans-11) and CLA, which is a favorable change in regard to current human dietary guidelines.