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Practical Aspects of Neuropathology: A Technical Guide for Working with the Nervous System
Andrew S. Fix
The Procter and Gamble Company, Miami Valley Laboratories, Cincinnati, Ohio 45253
Robert H. Garman
Consultants in Veterinary Pathology, Murrysville, Pennsylvania 15668
Toxicologic pathologists are evaluating tissues from the central and peripheral nervous systems with increasing frequency. This change is being driven by recently established regulatory guidelines and intense interest in developing pharmaceutical compounds to treat various nervous system disorders. However, morphologic evaluation of the nervous system by light or electron microscopy requires special understanding and effort. Here, we review the general concepts of fixation for the nervous system, explain perfusion procedures for optimal preservation, and provide information on handling tissues to avoid artifacts. In general, fixation with aldehydes is recommended for nervous tissue (a combination of paraformaldehyde and glutaraldehyde is preferred). Electron microscopic studies require fixatives of the highest purity possible, typically paraformaldehyde prepared fresh from powder mixed with high-grade glutaraldehyde. The final osmolality of the solution should be slightly hypertonic, in the range of 400-600 mOsmol. Slight hypertonicity is very important and will facilitate maintenance of vascular distention during whole-body perfusion, which is the best method for producing high-quality tissue preparations. Special effort is necessary for handling nervous tissue in a way that minimizes artifacts because chemical fixation is not completed immediately following the perfusion. These technical details should help toxicologic pathologists in their efforts to work with the nervous system, thereby increasing their effectiveness in supporting safety characterization of new test materials undergoing toxicologic assessments.
Key Words: Aldehydes • artifacts • fixation • immersion • immunohistochemistry • perfusion • special stains • tissue handling
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Toxicologic Pathology, Vol. 28, No. 1,
122-131 (2000)
DOI: 10.1177/019262330002800115
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