摘要
目的 测试Ki-67 MIB1克隆在甲状腺透明变梁状肿瘤(hyalinizing trabecular tumor,HTT)的免疫组织化学膜表达定位的最适染色条件,确认MIB1膜表达的客观方法学对照和可重复的染色标准.方法 纳入扁桃体、阑尾、肾脏、肝脏、肺、胰腺、皮肤、胎盘、前列腺、卵巢和羊膜各33例,对MIB1鼠抗人单克隆抗体行Envision两步法实验条件摸索.然后采用HTT和非HTT各16例以及17例硬化性肺细胞瘤(PSH)来观察各平台(Omnis、BenchMark Ultra、BondMAX)与手工检测对比MIB1的实际表达情况.结果 正常肺组织呈现弱的细胞膜表达,HTT为肿瘤细胞膜中等至强表达,而PSH为肿瘤细胞膜弱至中等表达.手工检测方式,高压热修复(121℃,2～2.5 min)或煮沸修复(100℃,20 min),MIB1效价浓度(1∶50～100)均能实现肺的细胞膜定位,与修复pH和所使用的二抗体系无明确关联.对比全自动平台发现,采用MIB1浓缩液1∶50稀释均无法实现正常肺的细胞膜表达.16例HTT MIB1膜染色,BenchMark Ultra和手工相比存在肿瘤细胞膜表达偏弱或无表达的明显差异,16例非HTT病例在各平台上均无细胞膜着色.17例PSH在3种全自动平台膜表达与手工方式相比,肿瘤细胞均存在膜表达不足或缺失的差异.结论 高压热修复或煮沸状态下,MIB1 1∶50～100,肺组织可以作为MIB1 HTT染色的理想外部对照.基于上述必备条件,可以实现近100％的HTT膜表达.
Abstract
Objective To test the optimal staining conditions for Ki-67 MIB1 clone in the immunohistochemical membrane ex-pression localization of hyalinizing trabecular tumor (HTT) of the thyroid,and to confirm the objective methodological control and re-producible staining standards for MIB1 membrane expression.Methods The samples of tonsils,appendix,kidney,liver,lung,pancreas,skin,placenta,prostate,ovary,and amniotic membrane,33 cases each,were included.Envision two-step experimental conditions for the MIB1 mouse anti-human monoclonal antibody were explored.Then,16 cases of HTT,16 cases of non-HTT and 17 cases of scle-rosing pulmonary cell tumors (PSH) were used to compare the actual MIB1 expression across different platforms (Omnis,BenchMark Ultra,BondMAX) and manual detection methods.Results Normal lung tissue exhibited weak cell membrane expression.HTT showed moderate to strong expression in tumor cell membranes,while PSH showed weak to moderate expression in tumor cell membranes.Manual detection methods with high-pressure heat retrieval (121°C,2-2.5 min) or boiling retrieval (100°C,20 min) at MIB1 concen-trations (1∶50-100) successfully localized to the lung cell membrane,with no clear correlation to the retrieval pH and the secondary an-tibody system used.Compared to fully automated platforms,MIB1 at a 1∶50 dilution failed to achieve membrane expression in normal lung tissue.In 16 cases of HTT,the membrane staining of MIB1 on BenchMark Ultra showed a significantly weaker or absent tumor cell membrane expression compared to manual methods.For 16 non-HTT cases,there was no membrane staining across all platforms.In 17 cases of PSH,tumor cells on all three fully automated platforms showed insufficient or absent membrane expression compared to manual methods.Conclusion With high-pressure heat retrieval or boiling,MIB1 at a concentration of 1∶50-100,lung tissue can serve as an ideal external control for MIB1 HTT staining.Based on these essential conditions,nearly 100％ membrane expression in HTT can be achieved.
维普
万方数据