Numerous methods used for the isolation of brain microvessels involve procedures which disturb the structural integrity of the cells and their organelles. In the present study, analysis of the adenylate energy charge and content as well as the incorporation of adenosine derivatives in isolated rat brain microvessels indicated a lesion of the mechanisms of energy production. The results show that experiments on isolated microvessels prepared by a mechanical homogenization exerting shear forces should be interpreted with caution when the rate of energy metabolism is a significant factor in the study.