A "recipe" for estimating neuron number in the mammalian spinal cord using modern stereological techniques
Department of Anatomy and Cell Biology, University of Bergen, N-5009 Bergen, Norway; Department of Pathology, The Central Hospital of Rogaland, N-4003 Stavanger, Norway
Department of Anatomy and Cell Biology, University of Bergen, N-5009 Bergen, Norway
Department of Anatomy and Cell Biology, University of Bergen, N-5009 Bergen, Norway
Stereological Research Laboratory, University Bartholin Building, DK-8000 Århus C, Denmark
Abstract
In this review article a brief introduction to the organisation and function of the mammalian spinal cord is given. This short account is followed by a detailed practical approach for estimating the number of neurons, glial and endothelial cells in the grey matter of the spinal cord. All steps, from vascular perfusion fixation, removal of the spinal cord from the spinal column, identification and isolation of individual spinal cord segments, embedding, sectioning, staining, and the finalstereological methods and statistical analysis are covered. Focus will be on the practical procedures, and the biological data obtained by these methods will only be discussed with respect to some methodological problems that have been encountered. The possibility of combining specific cell labelling techniques, such as tract tracing and immunocytochemistry, with stereological methods is also discussed, as these methods may solve some of the uncertainties and problems facing neuroscientists who want estimates of neuron number.