Image analysis software for automatic DNA ploidy assessment of archival solid tumours

Background: Image cytometry has proved to provide a good alternative to flo w cytometry for DNA ploidy measurement of archival tumors. However, when in teractively done this technique is unable to give statistically valuable re sults within an acceptable time for clinical oncology. Methods: An image cytometer was developed for fully automatic DNA ploidy qu antitation, focusing efforts on speed and accuracy. Software functionalitie s include systematic acquisition of fields on a microscopic slide, detectio n, localization and sorting of nuclei, computation of the DNA content toget her with post-processing tools, for a deeper analysis of the DNA ploidy dia gram. Results: DNA ploidy analysis of archival breast carcinoma samples illustrat es the accuracy of DNA ploidy measurements and the sensitivity in the detec tion of DNA ploidy abnormalities as a result of cell sorting. Conclusions: Fully automatic image cytometry is able to combine qualities o f flow cytometry (automatic analysis of a statistically significant collect ion of cell nuclei) with additional advantages: sorting of unwanted events (debris, stromal and inflammatory cell nuclei) and facilities for an a post eriori control of the quality of cell selection. This method is well suited to DNA ploidy analysis of archival cancer samples. Cytometry 37:267-274, 1 999. (C) 1999 Wiley-Liss, Inc.