胸腹水细胞蜡块制作方法的改良及应用

The improvement and application value of the preparation method for making cell blocks from pleural and ascitic fluid

目的 探讨不同固定方法下制作胸腹水细胞蜡块的效果及应用价值。方法 收集2022年6月至12月厦门大学附属第一医院病理科收检的胸腹水标本,选取经细胞病理学确诊见肿瘤细胞者48例,每份标本均平分为3份,离心后沉淀物分别按以下方法固定:A组10%中性缓冲甲醛固定4 h;B组薄层液基细胞学检查指定保存液固定4 h;C组加入4%中性甲醛固定液固定5 min,离心后沉淀物加入75%乙醇,1 min后吸弃75%乙醇,加入适量的95%乙醇,室温下静置塑形2 h;C组沉淀物较大者,沿纵轴每间隔3 mm平行切开,取其切面作为包埋面。3组分别制成细胞蜡块,比较蜡块制作成功率、蜡块肉眼观、蜡块切片HE染色效果、(肺腺癌)细胞蜡块切片免疫组织化学表达效果。结果 C组细胞蜡块制作成功率显著高于A组与B组。巨检:A组与B组固定后获得的沉淀物较松散、破碎;C组固定后获得的沉淀物聚集成块、富有弹性、结构完整、可见细胞层次,可更大限度的保存标本。细胞蜡块切片HE染色镜检:C组细胞量大,染色鲜艳,细胞轮廓清晰,细胞排列清楚可见,肿瘤细胞的核仁比较明显,优于A组与B组。C组细胞蜡块切片的CK7、napsin A、TTF-1 3种抗体的免疫组织化学表达效果均较好,阳性标记清晰易判,背景干净,优于A组与B组。结论 胸腹水细胞蜡块的制作,采用甲醛液固定后联合95%乙醇塑形2 h的效果较佳,细胞蜡块制作成功率高,获得的细胞蜡块完整,蜡块切片HE染色和免疫组织化学染色效果俱佳,可提高诊断准确率。

Objective Exploring the effects and application value of making pleural and ascitic fluid cell blocks with different fixation methods.Methods Collecting pleural and ascitic fluid specimens examined by the Department of Pathology at the First Affiliated Hospital of Xiamen University from June to December 2022,selecting 48 cases diagnosed with tumor cells by cytopathology.Each specimen was divided into three equal parts,and the sediment after centrifugation was fixed using the following methods:Group A was fixed with 10%neutral buffered formalin for 4 h;Group B was fixed with the designated preservative for ThinPrep cytologic examination for 4 h;Group C was fixed with 4%neutral formalin for 5 minutes,then after centrifugation,the sediment was added to 75%ethanol,left for 1 minute before discarding the ethanol,and then added to an adequate amount of 95%ethanol and left to set at room temperature for 2 h.For larger sediments in Group C,they were cut parallel along the longitudinal axis at 3 mm intervals,and the cut surface was used as the embedding surface.Cell blocks were made for all three groups,and the success rate of block making,gross appearance of blocks,HE staining effects of block sections,and immunohistochemical expression in cell block sections(for lung adenocarcinoma)were compared.Results The success rate of cell block preparation in Group C was significantly higher than in Groups A and B.Gross examination:the sediment obtained after fixation in Groups A and B was loose and fragmented;in Group C,the sediment formed cohesive,elastic blocks with intact structure and visible cellular layers,allowing for greater preservation of the specimen.Microscopic examination of HE-stained cell block sections:Group C had a larger quantity of cells with bright staining,clear cell outlines,and visible cell arrangement.The nucleoli of tumor cells were more evident in Group C,superior to Groups A and B.The immunohistochemical expression of CK7,napsin A,and TTF-1 antibodies in Group C cell block sections was better,with clear and easily interpretable positive markers,and a clean background,superior to Groups A and B.Conclusion The production of cell blocks from pleural and ascitic fluids shows better results when using formalin fixation followed by shaping with 95% ethanol for 2 h. This method yields a high success rate in cell block production, with the obtained cell blocks being intact. The HE staining and immunohistochemical staining of the cell block sections are both excellent, which can enhance diagnostic accuracy.