Fosså S D, Thorud E, Shoaib M C, Pettersen E O
Pathol Res Pract. 1986 May;181(2):200-5. doi: 10.1016/S0344-0338(86)80011-5.
Suspensions of single cell nuclei were obtained from 31 different samples of 7-9 years old paraffin-embedded bladder cancer biopsies. The DNA content of the ethidium-bromide-stained nuclei was analyzed by flow cytometry (FCM). The tumour stemline ploidy, as determined by FCM, was compared with that obtained by Feulgen scanning absorption cytometry (SACM) in imprints obtained from the same biopsy before it was fixed. Fifteen tumours that were diploid by FCM were also diploid by SACM. All of the 16 tumours with non-diploid DNA-stemlines by FCM were non-diploid by SACM, though minor differences between the ploidy values were occasionally seen when the results of the two methods were compared. The background activity due to cell debris was considerable and resulted in a mean variation coefficient (CV) of 5.1% for human spleen cells fixed and embedded before preparation for FCM in the same way as the tumour samples. In the tumour samples there was a large and unpredictable variation of the ratio between the DNA values of chicken erythrocytes (internal standard) and diploid cells. In some specimens this ratio would have resulted in incorrect evaluation of the FCM DNA histogram. The following method for evaluation of FCM histograms is therefore proposed: Single peak histograms obtained from paraffin embedded tissue should always be interpreted as representative for a diploid cell population. In FCM histograms from paraffin-embedded tissue with more than one peak, the first peak should be considered as representing diploid cells.
从31份7至9岁石蜡包埋膀胱癌活检组织的不同样本中获取单细胞悬液。通过流式细胞术（FCM）分析经溴化乙锭染色的细胞核的DNA含量。将FCM测定的肿瘤干系倍体与同一活检组织在固定前的印片中通过福尔根扫描吸收细胞术（SACM）获得的结果进行比较。15例FCM检测为二倍体的肿瘤，SACM检测也为二倍体。FCM检测DNA干系非二倍体的16例肿瘤，SACM检测均为非二倍体，不过比较两种方法的结果时，偶尔会发现倍体值存在细微差异。细胞碎片导致的背景活性相当高，对于与肿瘤样本相同方式固定和包埋后用于FCM检测的人脾细胞，平均变异系数（CV）为5.1%。在肿瘤样本中，鸡红细胞（内标）与二倍体细胞的DNA值之比存在很大且不可预测的差异。在某些标本中，该比例会导致对FCM DNA直方图的错误评估。因此，建议采用以下方法评估FCM直方图：从石蜡包埋组织获得的单峰直方图应始终解释为代表二倍体细胞群体。在来自石蜡包埋组织的FCM直方图中，若有多个峰，第一个峰应视为代表二倍体细胞。
