Estimation of the numerical densities of neurons and synapses in cerebral cortex

In this paper we discuss a stereological technique, 'the unfolding method', for a quantitative study of the nervous system [1,31]. Stereology implies a geometric analysis of structures and textures, and is a method to derive directly metric properties of structures from two-dimensional sections on the basis of geometrico-statistical reasoning [36,37]. Recent advances in the stereological method allow quantitative analysis [8,19,27,32]. Images on sections provide only two-dimensional information, but the stereological method can offer three-dimensional and quantitative information [19]. The need for quantitative analysis is more important and useful in the central nervous system (CNS) than in other organs. Two functional units, neurons and synapses, are of particular interest in evaluating CNS function. Numerical densities of neurons and synapses in rat visual cortex were estimated using the unfolding method at light and electron microscopic levels, respectively. Once the numerical densities of neurons and synapses were obtained, synapse-to-neuron ratios could be calculated. The ratios are interpreted as a means to obtain an index of interneuronal connectivity [9]. The unfolding method may become a powerful strategy in neuroscience research when numerical estimates are performed in restricted areas such as cortical layers II-IV, because this method is less time-consuming than other stereological methods [6,21,22].