The objective of this study was to evaluate the effects of adding flaxseed oil (FO) to feed on the incorporation of n-3 PUFA in tilapia heads. Tilapia were given diets with increasing levels of FO (0.00, 1.25, 2.50, 3.75 and 5.00% for treatments A, B, C, D and E, respectively), as a source of LNA for 150 days. The proximate composition of the heads indicated high nutritional value and 40 FA (fatty acids) common to all treatments were identified in total lipids. Intake of LNA caused storage of LNA and sequential desaturation-elongation to eicosapentaenoic acid (EPA) and DHA. With increasing levels of FO in the diet, the content of LNA in tilapia heads increased (1.7 and 14.0% for diets A and E, respectively), as well as the contents of EPA (0.1 and 0.9% for diets A and E, respectively) and DHA (0.5 and 1.8% for diets A and E, respectively). Adding FO to tilapia feed markedly increased the total content of n-3 PUFA (3.0 and 21.1% for diets A and E, respectively), decreased the total content of n-6 PUFA (41.3 and 24.9% for diets A and E, respectively), and consequently resulted in a decrease in the n-6/n-3 ratio (13.8 and 1.2 for diets A and E, respectively). Therefore, feeding tilapia with FO is a good way of valorizing this part of the fish by creating a valuable nutritional food source.