Estimators of the precision of stereological estimates: An example based on the CA1 pyramidal cell layer of rats
Section snippets
Animals and tissue processing
Two male Wistar rats (305 and 315g) were deeply anesthetized with sodium pentobarbital and perfused transcardially with ∼500ml of a solution of 2% paraformaldehyde and 1% glutaraldehyde in 0.15M phosphate buffer (pH 7.4). The brains were removed immediately after perfusion and postfixed for 6 h. Subsequently, the brains were divided into left and right hemispheres and non-telencephalic structures were removed. The dorsal cortex was removed by a cut in the horizontal plane above the hippocampal
Volume distribution
The volume distributions of the CA1 pyramidal cell layer in a horizontal and a coronal series of sections, are illustrated in Fig. 1a and b respectively. In Fig. 1a, several sharp peaks are observed in the first one-third of the horizontal series. These peaks correspond to sections that cut the CA1 pyramidal cell layer in a plane that is close to the plane of the layer itself, i.e. in a plane that is close to being tangential to the CA1 pyramidal cell layer. These include sections in which the
Discussion
The main focus of this study has been to describe the issues that should be addressed when making a proper decision about the precision of a stereological estimation procedure, the CE, needed for a specific study. This has been done by collecting data from relatively large numbers of sections and sample sites (i.e. over-sampling) in an irregularly shaped brain structure, CA1 of the rat hippocampus. We then calculated the precision of the estimates for the different combinations of subsets of
Acknowledgments
We gratefully acknowledge the technical assistance of Irmgard Amrein. We also want to thank the participants of several NeuroStereology Workshops for the inspiration that prompted us to examine the CE in more detail. This work was supported by Swiss National Foundation (SNF) and NCCR ‘Neural Plasticity & Repair,’ and The Danish Health Sciences Research Council.
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