Laetiporus sulphureus is a species of bracket fungus found mainly in Europe and North America. Its common names are sulphur polypore, sulphur shelf, and chicken of the woods because of its taste which may also resemble the taste of crab or lobster. Its fruit bodies grow as golden-yellow shelf-like structures on tree trunks and branches. Like other bracket fungi, they may last many years and fade to pale grey or brown. The under surface of the fruit body is made up of tubelike pores. Laetiporus sulphureus is a saprophyte and causes brown cubical rot in the heartwood of trees on which it grows. Unlike many bracket fungi, it is edible when young. In this study, the lipid profile of the fruiting bodies of L. sulphureus has been studied using GC-FID and Iatroscan TLC FID methodologies. Different types of extractions including high energy techniques were employed to identify the optimum conditions for higher yield of lipid isolation. Optimum extraction methodologies provided two fractions, one containing neutral and polar lipids (TGs, sterols, GL, phospholipids) and the other mainly attributed to fungal carotenoids and pigments. The existence of relatively high amount of sterols may be correlated to fungus pharmaceutical properties. Fatty acid analysis indicated a predominant level of polyunsaturated fatty acids (PUFA) followed by saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA).