Preparation and analysis of the central nervous system

For many pathologists, neuropathology is intimidating. Practical approaches for nervous tissue histologic evaluations to meet both routine and advanced study designs can lead to rewarding neuropathology efforts. Cost-effective, high-quality histologic evaluations can occur if animals are exsanguinated quickly, brains removed carefully to maintain structural integrity and avoid dark neuron artifact, immersion-fixed quickly and thoroughly, and trimmed and processed to consistently survey multiple areas. While brightfield examination of H&E-stained sections is generally sufficient for survey evaluations, epifluorescent assessment of neuronal autofluorescence facilitates recognition of neurodegeneration in H&E-stained sections. Fluoro-Jade B or specialized immunohistochemical stains may be required to answer specific questions. Evaluations require that both technical staff and pathologists have a working knowledge of a few easily identified neuroanatomic landmarks and familiarity with use of a detailed brain atlas. At least four coronal sections should be routinely surveyed from young adult rats, with evaluation of comparable areas in other laboratory animal species. This number should be at least doubled if there is reason to suspect morphologic changes in the CNS. This article focuses on technical details of efficient specimen preparation for neuropathologic evaluations involving relatively large numbers of rodents, as well as a practical approach to basic neuroanatomic site identification.