R A Williams1, J P Baak, G A Meijer, I G Charlton. 1. St. Vincent's Department of Anatomic Pathology, Wangaratta and District Pathology Service, University of Melbourne, Australia.
Abstract
OBJECTIVE: To analyze the influence of measuring parameters on the quality of DNA histograms, and to compare the results of DNA image cytometry (ICM) and flow cytometry (FCM). STUDY DESIGN: Material from 20 breast cancer cases was analyzed. The protocol for ICM analyzed (1) measuring system and image analysis parameters, (2) the quality of fixation, and (3) the presence of cytoplasm around nuclei. In Feulgen-stained slides (both fresh imprints and cytocentrifuge preparations) at least 200 nuclei were examined using a commercially available ICM system. DNA FCM on disaggregates was performed, also. RESULTS: Strict adherence to the protocol gave reliable DNA ploidy profile results. In contrast, deviations from the protocol gave very broad histograms. With the protocol, similar results were found for imprints and cytocentrifuge samples. Duplicate "blind" assessments were highly reproducible. DNA indices measured by FCM and ICM (on cytocentrifuge samples) correlated highly as well. CONCLUSION: With the protocol used, DNA ploidy profiles obtained by ICM from fresh imprints and cytocentrifuge disaggregate cell preparations were comparable.
OBJECTIVE: To analyze the influence of measuring parameters on the quality of DNA histograms, and to compare the results of DNA image cytometry (ICM) and flow cytometry (FCM). STUDY DESIGN: Material from 20 breast cancer cases was analyzed. The protocol for ICM analyzed (1) measuring system and image analysis parameters, (2) the quality of fixation, and (3) the presence of cytoplasm around nuclei. In Feulgen-stained slides (both fresh imprints and cytocentrifuge preparations) at least 200 nuclei were examined using a commercially available ICM system. DNA FCM on disaggregates was performed, also. RESULTS: Strict adherence to the protocol gave reliable DNA ploidy profile results. In contrast, deviations from the protocol gave very broad histograms. With the protocol, similar results were found for imprints and cytocentrifuge samples. Duplicate "blind" assessments were highly reproducible. DNA indices measured by FCM and ICM (on cytocentrifuge samples) correlated highly as well. CONCLUSION: With the protocol used, DNA ploidy profiles obtained by ICM from fresh imprints and cytocentrifuge disaggregate cell preparations were comparable.