Analysis of measuring system parameters that influence reproducibility of morphometric assessments with a graphic tablet.

ABSTRACT

The morphometric analysis of nuclear characteristics by means of a graphic tablet is, in principle, objective and highly reproducible. However, a recent study found considerable variation in the morphometric assessments, which was in contrast to the findings of others. The way in which measurements were performed differed in these studies. Therefore, measuring system factors that can potentially influence the quantitative results were analyzed systematically. One observer, experienced in microscopic analysis and working with a commercially available graphic tablet, conducted all the measurements, thus excluding interobserver variation. The tracing speed, localization (on the graphic tablet), magnification, pen and cursor usage, shape, and orientation on the graphic tablet were analyzed. A nomogram was developed for cursor application that indicates the relation between "projected" particle size, tracing speed, and required coefficient of variation (CV). When the influence of these factors is taken into account, a measuring system can be tuned optimally. With such a regimen, the CV can be kept below 1.5%. Our results show that in the assessment of morphometric features with the use of a graphic tablet, errors due to the measuring system can be virtually eliminated.