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The saucor, a new stereological tool for analysing the spatial distributions of cells, exemplified by human neocortical neurons and glial cells

by Anette K. Stark, Hans-Jørgen G. Gundersen, Jonathan Gardi, Bente Pakkenberg and Ute Hahn
Thiele Research Reports Number 13 (November 2009)

The three dimensional spatial arrangement of particles or cells, for example glial cells, with respect to other particles or cells, for example neurons, can be characterized by the radial number density function, which expresses the number density of so called "secondary" particles as a function of their distance to a "primary" particle.

The present paper introduces a new stereological method, the saucor, for estimating the radial number density from thick isotropic uniform random (IUR) or vertical uniform random (VUR) sections. In the first estimation step, primary particles are registered in a disector. Subsequently, smaller counting windows are drawn with random orientation around every primary particle, and the positions of all secondary particles within the windows are recorded. The shape of the counting windows is designed such that a large portion of the volume close to the primary particle is examined and a smaller portion of the volume as the distance to the primary object increases. The experimenter can determine the relation between these volumina as a function of the distance by adjusting the parameters of the window graph, and thus reach a good balance between workload and obtained information. Estimation formulae based on the Horvitz-Thompson theorem are derived for both IUR and VUR designs.

The method is illustrated with an example where the radial number density of neurons and glial cells around neurons in the human neocortex is estimated using thick vertical sections for light microscopy. The results indicate that the glial cells are clustered around the neurons and the neurons have a tendency towards repulsion from each other.

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