Preparation of Brain Slices for Macroscopic Study by the Copper Sulfate-Phenol-Ferrocyanide Technique

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Abstract
The technique of Mulligan (J. Anat., 65: 468–72, 1931) for preparing gross brain slices has been modified to include improvements in fixation, slicing and staining. The iron alum-tannic acid steps have been eliminated and potassium ferrocyanide substituted. Washing times have been reduced and a detailed routine for fixation and slicing developed. After postmortem removal, human brains were perfused with 40% formalin and kept suspended in 10% formalin for 2–4 wk. Serial slices, 4 mm thick, were cut with an electrically driven rotary-blade meat slicer, washed in running water for 12 hr, immersed in CuSO4-phenol solution (0.5%: 5%) at 60 C for 6 min, washed again in running water for 5 min, and then treated with a 2% solution of potassium ferrocyanide until a red-brown color developed in the gray matter, usually within 30–60 sec. The slices were then washed again for 5 min and stored in 10% formalin. Brains of patients over 60 yr of age frequently stained poorly and were not used.