AbstractCerebellar and cerebral cortices were frozen under various conditions; within 30 seconds of circulatory arrest, after eight minutes of asphyxiation, after ten minutes of perfusion with glutaraldehyde and after perfusion with this fixative and osmium tetroxide postfixation. Ethanol was used as the solvent in freeze substitution of these tissues. The resulting EMs closely resembled those of similar material freeze substituted in acetone. There were no differences in extracellular space even though differences have been reported between the space in EMs of conventionally fixed material dehydrated with ethanol or acetone.