The effect of improper realignment of serial sections on surface area measurements of 3D-reconstructed objects
Abstract
Stereological estimation of surface area of three-dimensional objects using serial sections usually requires perfect realignment of the sections. In physical sectioning often fiducial markers are used to realign the sections. Nevertheless shrinking and other deformations of the slices frequently make it impossible to obtain a precise reconstruction of the spatial arrangement. This inaccuracy causes an error in surface area estimation using methods as e.g. the spatial grid. As shown in the paper, it leads to an overestimation of the surface area. The error is related to the deviation of the “true” slice positions and a simple method is suggested to calculate an upper bound of the error by comparing the relative positions of the fiducial marker profiles on the sections. This method is successfully applied to the estimation of a fish brain’s surface area.