A simple way of quantifying immunostained cell nuclei on the whole histologic section

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Abstract
Background Quantification of nuclei, immunostained for proliferation or differentiation markers, is widely recommended for prognostication and treatment of numerous solid tumors. However, many problems concerning reproducibility of the results obtained still remain, partially due to the lack of standardization of the method used and to the difficulty of taking into account tumor tissue heterogeneity. Technical solutions exist but they are expensive and time consuming, and their complexity limits their diffusion in routine pathology departments. Methods In the present study, we use a simple, fast and inexpensive system of acquisition, a 4,000 dots‐per‐inch slide scanner, to get a single true color numerical image of the whole histologic section. The analysis is done thanks to fully automatic image processing, allowing computation of the stain proportion on the whole histologic section, as well as in “hot spots.” Results After having precisely fixed settings of the slide scanner, one can easily detect counterstained and immunostained structures, by image analysis. This simple and stable processing can be done under 6 min. Conclusions The association of immunohistochemistry, stable acquisition device and computer‐assisted image analysis provides an objective, reproducible, and easy to standardize quantification of the nuclear markers, in relation to the total tumor tissue architecture. Cytometry Part A 56A:37–45, 2003.