Torp S H, Johannesen E, Lindboe C F
Department of Pathology, Institute of Morphology, University Hospital of Trondheim, N-7006 Trondheim, Norway.
Clin Mol Pathol. 1995 Aug;48(4):M191-3. doi: 10.1136/mp.48.4.m191.
Aims-To compare immunostaining between the original Ki67 monoclonal antibody and a new polyclonal Ki67 antibody on frozen and paraffin wax sections of human glioblastomas.Methods-Frozen sections and formalin fixed, paraffin wax embedded sections of the same tumour specimens were included in the study (10 cases). Half of the paraffin wax sections were pretreated in a microwave oven. Standard immunohistochemical techniques were used (avidinbiotin peroxidase complex). Five high power fields were examined using an eye-piece graticule, and 500 to 2000 tumour cells were counted. The labelling index was defined as the percentage of positive tumour cells.Results-The Ki67 monoclonal antibody displayed positive immunostaining in all frozen sections (median labelling index 5.9, range 2.6-11.4) whereas only four paraffin wax sections stained positively and only after pretreatment in a microwave oven. The polyclonal Ki67 antibody elicited positive staining in both frozen sections (median labelling index 13.7, range 6.7-21.5) and in paraffin wax sections (median labelling index 12.0, range 2.2-22.7) but only after pretreatment in a microwave oven.Conclusion-The Ki67 monoclonal antibody is not recommended for use on paraffin wax sections of glioma tissues whereas the new polyclonal Ki67 antibody provides satisfactory immunostaining on both frozen and paraffin wax sections.
目的——比较原始的Ki67单克隆抗体与一种新的Ki67多克隆抗体在人胶质母细胞瘤冰冻切片和石蜡切片上的免疫染色情况。方法——本研究纳入了同一肿瘤标本的冰冻切片以及经福尔马林固定、石蜡包埋的切片(10例)。一半的石蜡切片在微波炉中进行预处理。采用标准免疫组织化学技术(抗生物素蛋白-生物素过氧化物酶复合物法)。使用目镜测微尺检查5个高倍视野,计数500至2000个肿瘤细胞。标记指数定义为阳性肿瘤细胞的百分比。结果——Ki67单克隆抗体在所有冰冻切片上均显示阳性免疫染色(标记指数中位数为5.9,范围2.6 - 11.4),而只有4张石蜡切片呈阳性染色,且仅在微波炉预处理后。多克隆Ki67抗体在冰冻切片(标记指数中位数为13.7,范围6.7 - 21.5)和石蜡切片(标记指数中位数为12.0,范围2.2 - 22.7)上均引发阳性染色,但也仅在微波炉预处理后。结论——不推荐Ki67单克隆抗体用于胶质瘤组织的石蜡切片,而新的多克隆Ki67抗体在冰冻切片和石蜡切片上均能提供满意的免疫染色。