Intestinal absorption of [SH]octanoic acid and [SH]decanoic acid was investigated in the rat by electron microscope autoradiography, The common duct (bile and pancreatic common duct) of the rats was diverted and a loop of the duodenum was cannulated 24 h later. The lipid mixture to be investigated was introduced into each experimental loop, and after 15 rain or less the loop was removed. One part of each loop was used to determine the distribution of radioactivity in different lipid fractions, and an autoradiographic study was performed on the other part of the loop.Radioactivity distribution studies confirmed that medium chain fatty acids are absorbed in their nonesterified form and established that these fatty acids are absorbed much more rapidly than oleic acid. Autoradiographic studies indicated that the medium chain fatty acids are taken up in a molecular or aggregate molecular form, leave the epithelial cells by way of the lateral plasma membrane, and are next found in the blood capillaries. Our results suggest that the Golgi complex does not play an important role in the absorption of unesterified fatty acids.It has been shown that the absorption of fatty acids, which occurs in the upper part of the small intestine, differs according to the chain length of the fatty acids. Long chain fatty acids are esterified into triglycerides and delivered to the lymph in the form of chylomicrons. Short chain fatty acids are transported by the portal blood as free fatty acids bound to albumin. Fatty acids of intermediate chain length are transported by both the lymph and the blood, in a ratio related to the number of carbon atoms (3,4,5,13, 19).The different pathways of fatty acid absorption suggest the involvement of different subcellular components. Electron microscopy has been used by several groups (7,17,18,25,36) to study the structures involved in the intestinal absorption of fats and particularly of long chain fatty acids. These studies, together with biochemical investigations, demonstrated that the apical vesicles of the absorptive cell are the site of triglyceride resynthesis (28,29,30,31). Electron microscope autoradiography indicated that long chain fatty acids are taken up from a molecular or micellar dispersion (34,35). Studies by Dermer (15) and from our laboratory (8, 11) have shown that long chain fatty acids enter the cell by simple diffusion. These studies have especially confirmed that the endoplasmic reticulum and the Golgi apparatus are involved in the intracellular transport of long chain fatty acids.