Despite numerous studies, the importance which the tissue or the composition of the diet may have in the biological distribution of each fatty acid is not well known. To determine the importance of tissue origin and dietary fatty acids in the fatty acid composition of cell phospholipids, 54 male adult rats were fed isocaloric diets for one month varying only in their fatty acid compositions. The fat component of the six experimental groups was derived from olive oil, sunflower oil, fish oil, soybean oil, palmitic acid, or 82% palmitic acid plus 18% soybean oil, supplying the essential fatty acid. The fatty acid composition of phospholipids from thymocytes, pancreatic exocrine, muscle and adipose tissues was studied by gas-chromatography. The tissue of origin was a more important source of variation than diet in the fatty acid content of the cell phospholipids except for palmitic acid (16:0), eicosapentaenoic acid (20:5 n-3), and docosahexaenoic acid (22:6 n-3). This study points out the complexity of the interrelations between different families of fatty acids and of the specificity of each tissue to changes in the composition of dietary fatty acids, as well as the inconvenience of speaking from the dietary point of view of groups of fatty acid families based on the position of the double bond, since their individual behaviour, including saturated fatty acids, is very different in the face of dietary manipulation. The study also highlights the different behaviour of each of the fatty acids in relation to the others in the diet in each of the tissues, a circumstance which should be taken into account when evaluating the biological effects in both epidemiological and experimental studies.