Microvessels were isolated from autopsied human brain using a simple procedure involving disruption, sieving, and centrifugation on a sucrose density gradient. The present procedure is characterized by isolation, from frozen autopsied brain, of materials either from the cerebral cortex or white matter, and subsequent separation of the capillary fraction from the large vessel fraction. The preparation appears highly purified under phase-contrast microscopic examination. The purity was also established by the enrichment of gamma-glutamyl transpeptidase activity and by the nearly negligible cerebroside content in the vessel fractions as compared to the brain homogenate.