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This study aimed to evaluate the potential of the marine microalgae Nannochloropsis oceanica as a sustainable source of n-3 polyunsaturated fatty acids (n-3 PUFA) for hen eggs enrichment. During 4 weeks, hens were fed with 3% (w/w) of Nannochloropsis oceanica supplemented diet. Throughout the assay, eggs were analyzed according to several nutritional and physical parameters, namely: (i) protein, fat, and ash content; (ii) fatty acid profile; (iii) thickness and colour of the shell; (iv) total egg weight; (v) protein quality (HU) and (vi) yolk colour. A remarkable increase in eicosapentaenoic (EPA), from 2.1 ± 0.1 to 5.2 ± 1.2 mg/100 g, and docosahexaenoic (DHA), from 50.3 ± 4.0 to 105 ± 18 mg/100 g, fatty acids was observed. Yolk colour also changed significantly according to the La Roche scale, from 9.6 ± 0.8 to 11.4 ± 0.8 (more orange). Feed supplementation did not lead to changes in the remaining analyzed parameters. A shelf life study, carried out for 28 days at room temperature, showed a decrease in eggs protein quality. In conclusion, eggs from hens fed with Nannochloropsis oceanica had a yolk colour more appealing to consumers and higher levels of EPA and DHA, allowing its classification as high in n-3 PUFA (CE nº 1924/2006).