Digital stereology in neuropathology

Two-dimensional quantitative methods have frequently been used to address questions in neuropathological research; however, they face important limitations which design-based stereology may overcome by offering a set of methods to quantify two-dimensional histological sections into three-dimensional structural knowledge. Accordingly, stereology is a science based on statistical sampling principles and geometric measures. The application of stereology to neuropathological studies allows the researcher to efficiently obtain a precise estimate of various structural quantities. This neuropathological review will therefore present the relevant stereological estimators for obtaining reliable quantitative structural data from brains and peripheral nerves when using digital light microscopy. It is discussed how to obtain brain and nerve fibre samples to fulfil the requirements for the estimators. A presentation of design-based stereological probes for obtaining global volume, total number, local volume, total length, total surface area, cross-sectional area and diameter will be followed by discussion of the error variances of the methods. No mathematical equations or calculations are shown in this review.