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Abstract
A method of perfusion-fixation of the human brain is described and compared with immersion-fixation
by immunoperoxidase staining for several substances (tyrosine hydroxylase, substance
P, choline acetyltransferase, glutamate decarboxylase, Met-enkephalin, and neuron-specific
enolase) in human striatum. Results from 1-cm slices fixed by immersion for 1, 2,
4 and 8 days were compared with results from slices of perfused brain postfixed for
the same time periods. The fixative used in all steps was 4% paraformaldehyde at 4
degrees C. In the immersion-fixed brains, optimal immunoreaction for tyrosine hydroxylase
and glutamate decarboxylase was limited to a depth of 1-2 mm from the surface of the
brain slice. In contrast, staining density in perfusion-fixed brains was relatively
homogeneous and of high quality. The other antigens studied displayed more uniform
staining throughout the section with both perfused and immersed brains. Investigators
intending to study human brain immunohistochemistry using immersion-fixation should
be aware of the possibility of depth-related variations in staining intensity and
would be wise to determine whether this effect is significant for the antigens they
choose to study.