Lost caps in histological counting methods.

BACKGROUND: In methods with the goal of counting objects in a sectioned tissue volume by examining their profiles or segments in the sections, lost caps, i.e., small object fragments unnoticed or missing at the section surfaces, are an unavoidable issue. METHODS AND RESULTS: The problem of lost caps is examined as it applies to four methods for counting in histological sections, the method usually referred to as the Abercrombie correction, the empirical method, the optical disector, and the physical disector. CONCLUSIONS: Lost caps are an insoluble problem in the Abercrombie method; the lost caps error correction factor should be incorporated into the Abercrombie equation. Lost caps cancel out in the optical disector. The empirical method logically requires, to avoid lost caps error, either a preliminary blind identification of object segments in the serial sections or identification of segments with reference to adjacent sections in the counting sections. Similarly, the physical disector method requires either a preliminary blind identification of object segments in both look-up and sampling sections, or use of three sections rather than two.