Risberg B, Stål O, Bjelkenkrantz K, Hatschek T, Franzén G, Arvidsson S, Nordenskjöld B
Acta Radiol Oncol. 1985 Nov-Dec;24(6):537-44. doi: 10.3109/02841868509134428.
Sections with a thickness of 50 micron from 34 formalin fixed, and paraffin embedded tumours were deparaffinized and hydrated and the cells were disaggregated with pepsin. The cells were stained with Hoechst 33258 and a rapid method for static fluorometry was used. DNA ploidy and the fraction of cells in S-phase were estimated and compared with results obtained with freshly prepared cells. The coefficients of variation for the tumour stemlines for both the fresh and the embedded material were approximately 6 per cent. There was a good correlation between DNA ploidy determined from fresh and embedded material, respectively; there was also a close correlation between the fractions of S-phase cells estimated from fresh and embedded tumours. It was also possible to cut out small histologically defined lesions, such as carcinoma in situ of the breast from thick paraffin sections and obtain DNA histograms with high resolution.
从34个经福尔马林固定并石蜡包埋的肿瘤中切取厚度为50微米的切片，进行脱石蜡和水化处理，然后用胃蛋白酶将细胞解离。细胞用Hoechst 33258染色，并采用快速静态荧光测定法。估计DNA倍性和S期细胞分数，并与用新鲜制备的细胞获得的结果进行比较。新鲜材料和包埋材料的肿瘤干细胞系变异系数约为6%。分别由新鲜材料和包埋材料测定的DNA倍性之间有良好的相关性；从新鲜肿瘤和包埋肿瘤估计的S期细胞分数之间也有密切相关性。还可以从厚石蜡切片中切出小的组织学定义的病变，如乳腺原位癌，并获得高分辨率的DNA直方图。
