The interaction between magnesium (Mg) and tris(8-hydroxy-quinoline) aluminum (Alq3) has been studied using high-resolution electron energy-loss spectroscopy (HREELS). It was found that deposition of magnesium on the Alq3 film gave rise to clear changes in the HREELS spectra. The changes are attributed to the weakly bounded Mg atoms on the Alq3 layer. Interestingly, for a given amount of magnesium (Mg to Al atoms ratio=3) on Alq3 film, remarkable changes were observed in the HREELS spectra when the sample was heated. A loss peak at 81meV, which was assigned to Mg-O stretch mode, appeared upon annealing and increased in intensity as the annealing temperature increased up to about 360K. This suggested that the diffusion of Mg atoms into the Alq3 layer and the reaction between Mg and Alq3 molecule occurred at the temperature range investigated. The present work has provided direct evidence for the strong interaction between magnesium and Alq3.