Previously, all stereological estimation of particle number and sizes were based on models and notoriously gave biased results, were very inefficient to use and difficult to justify. For all references to old methods and a direct comparison with unbiased methods see recent reviews.The publication in 1984 of the DISECTOR, the first unbiased stereological probe for sampling and counting 3—D objects irrespective of their size and shape, signalled the new era in stereology — and give rise to a number of remarkably simple and efficient techniques based on its distinct property: It is the only known way to obtain an unbiased sample of 3-D objects (cells, organelles, etc). The principle is simple: within a 2-D unbiased frame count or sample only cells which are not hit by a parallel plane at a known, small distance h.The area of the frame and h must be known, which might sometimes in itself be a problem, albeit usually a small one. A more severe problem may arise because these constants are known at the scale of the fixed, embedded and sectioned tissue which is often shrunken considerably.
Unbiased and Minimum-Variance Unbiased Estimation of Estimable Functions for Fixed Linear Models with Arbitrary Covariance Structure