Williams R A, Baak J P, Meijer G A, Charlton I G
St. Vincent's Department of Anatomic Pathology, Wangaratta and District Pathology Service, University of Melbourne, Australia.
Anal Quant Cytol Histol. 1996 Dec;18(6):429-37.
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).
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.
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.
With the protocol used, DNA ploidy profiles obtained by ICM from fresh imprints and cytocentrifuge disaggregate cell preparations were comparable.
分析测量参数对DNA直方图质量的影响，并比较DNA图像细胞术（ICM）和流式细胞术（FCM）的结果。
分析了20例乳腺癌病例的材料。ICM分析方案包括（1）测量系统和图像分析参数，（2）固定质量，以及（3）细胞核周围细胞质的存在情况。在福尔根染色的载玻片（新鲜印片和细胞离心涂片）上，使用市售的ICM系统检查至少200个细胞核。同时也对解离后的样本进行了DNA流式细胞术检测。
严格遵守方案可得出可靠的DNA倍性分析结果。相反，偏离方案会得到非常宽泛的直方图。按照方案，印片和细胞离心涂片样本的结果相似。重复的“盲法”评估具有高度可重复性。FCM和ICM（对细胞离心涂片样本）测量的DNA指数也高度相关。
使用该方案，ICM从新鲜印片和细胞离心涂片解离细胞制备物中获得的DNA倍性分析结果具有可比性。
