Immunohistochemistry of neural markers for the study of the laminar architecture in celloidin sections from the human cerebral cortex

Morphometric studies of the cerebral cortex in celloidin sections provide r eliable quantitative estimates of cytoarchitectural features in individual brain regions. To increase our knowledge about the morphology and distribut ion of neuronal and glial cell types using specific cellular markers, we co mpared two methods of celloidin removal/antigen recovery, and subsequent im munohistochemical staining of free floating sections with specific antibodi es. The method based on methanol and NaOH for celloidin removal was the mos t adequate for optimal recovery of immunoreactivity of the neural markers N F200, MAP2, GFAP, calretinin, parvalbumin, calbindin-D-28kD, and synaptophy sin. The other method, based on a treatment with ethanol/ether and formic a cid, gave good results in the immunostaining of NF200, GFAP and MAP2, but n ot the other markers named above. The immunostained sections were compared with nearby sections stained with cresyl violet in order to assign the immu noreactive structures to individual layers in the prefrontal cortex. Sectio ns from blocks not embedded in celloidin showed a comparable distribution o f all the antigens included in the present study. The present paper provide s an antigen recovery technique for celloidin sections that can be applied to optimize studies on the cytoarchitecture and distribution of specific ne ural elements in the human cerebral cortex. (C) 1999 Elsevier Science B.V. All rights reserved.