Identification of a potential artifact in the use of electron microscope autoradiography to localize saturated phospholipids in cells.

The suitability of electron microscope autoradiography for sutdying the uptake and intracellular localization of lipid vesicles (liposomes) containing radiolabeled saturated phospholipids has been examined. Data are presented showing that preparation of specimens for electron microscope autoradiography by conventional methods is accompanied by significant translocation and intercellular redistribution of radiolabeled saturated lipids, causing spurious labeling patterns. Intercellular redistribution of radiolabeled lipid was demonstrated by mixing glutaraldehyde-fixed mous L1210 cells that had been incubated with sonicated lipid vesicles containing [H] dipalmitoyl phosphatidylcholine with an indicator cell population (fixed avian erythrocytes) which had not been exposed to vesicles and showing that after electron microscope processing radiolabeled grains were present in both cell types. The same redistribution artifact also probably affects the intracellular localization of radiolabeled lipids. This artifact is discussed in relation to previous work in which autoradiographic methods have been used for ultrastructural localization saturated phospholipids in cells and tissues.