When laying hen diets are enriched with omega-3 polyunsaturated fatty acids to generate value-added eggs for human consumption markets, concentrations of alpha-linolenic (ALA), eicosapentaenoic (EPA), and docosahexaenoic acids (DHA) in the yolk can reach 250 mg/50 g whole egg. Flaxseed, a rich source of ALA, is commonly used for omega-3 enrichment; however, the impact of dietary flaxseed source (extracted oil vs. milled seed) on fatty acid transfer to egg yolk in laying hens is unknown. Therefore, transfer of ALA, EPA, and DHA into egg yolk from extracted flaxseed oil or milled flaxseed was evaluated in Hy-Line W-36 laying hens over an 8-week feeding period (25 to 33 wk old). Hens (n = 132) were randomly housed with 3 birds/cage (4 replicates/treatment) for each of the 11 treatment groups. Diets were isocaloric and consisted of a control diet, 5 flaxseed oil diets (0.5, 1.0, 2.0, 3.0, or 5.0% flaxseed oil), and 5 milled flaxseed diets (calculated flaxseed oil concentration from milled flaxseed 0.5, 1.0, 2.0, 3.0, 5.0%). Increasing dietary concentrations of flaxseed oil and milled flaxseed resulted in increased ALA, EPA, and DHA concentration in egg yolk, and fatty acid deposition from flaxseed oil was 2 times greater compared to milled flaxseed when fed at the same dietary inclusions (P < 0.01). Egg yolk EPA and DHA concentrations were not different due to oil or milled source (P = 0.21); however, increasing dietary inclusion rates of flaxseed oil from either source increased yolk EPA and DHA (P < 0.01). Hens fed either flaxseed oil or milled flaxseed resulted in reduced BW change as dietary concentrations increased (P = 0.02). Feed efficiency increased as flaxseed oil increased in concentration, while feeding milled flaxseed decreased feed efficiency (P = 0.01). Analysis of the nitrogen corrected apparent metabolizable energy of flaxseed oil resulted in 7,488 kcal/kg on an as-fed basis. Dietary flaxseed oil improved feed efficiency and increased ALA deposition into yolk compared to a milled source, demonstrating flaxseed oil to be a viable alternative for ALA egg enrichment.