Objective: The aim of this study was to investigate the effects of simultaneous supplementation of laying hens with alpha-linolenic acid (ALA) resources (flax, perilla, and Eucommia ulmoides [E. ulmoides] seeds) and eicosapentaenoic acid/docosahexaenoic acid (EPA/DHA) resources (Schizochytrium sp.) on egg quality and n-3 polyunsaturated fatty acids (PUPA) profile. Methods: Dietary treatments were as follows: i) diet C (control diet); ii) diet F (diet C+10% flaxseeds); iii) diet P, (diet C+10% perilla seeds); iv) diet E (diet C+10% E. ulmoides seeds); v) diet A (diet C+1.5% microalage); vi) diet AF (diet C+10% flaxseeds+1.5% microalage); vii) diet AP (diet C+10% perilla seeds+1.5% microalgae); viii) diet AL (diet C+10% E. ulmoides seeds+ 1.5% microalage). Results: Egg weight, yolk weight and production ratio were not significantly affected by either algae or in combination with seeds (p>0.05). No significant difference was observed in ALA and DHA concentration in eggs between thawed, perila, and E. ulmodies seeds supplementation alone (p>0,05), N-3 PUPA in eggs was slightly improved by microalgae supplementation. The best supplementation, a combination of microalgae and perilla seeds, elevated (p<0.05) ALA from 19,7 to 202.5 mg/egg and EPA+DHA from 27.5 to 159.7 mg/egg. Highest n-3 PUPA enrichment (379.6 mg/yolk) was observed with supplementation of a combination of perilla seed and microalgae (362.2 mg/yolk), followed by a combination of flaxseed and microalgae (348.4 mg/yolk). The ALA, EPA, and DHA content obtained with a combination of microalgae and seeds surpassed the total sum of that obtained with microalgae or ALA-seeds alone. Conclusion: It is feasible to enrich eggs with n-3 PUFAs by perilla or E. ulmodies seeds instead of flaxseeds. Simultaneous supplementation of microalgae and seeds helped improve the transfer from EPA and docosapentaenoic acid into DHA.