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Masson三色染色常见问题分析与质量控制
周颖颖1,冼志红1*,张顺民2,高名士3,虞文伟4,宿杰·,阿克苏5
0
(1. 海军军医大学(第二军医大学)第三附属医院病理科, 上海 200438;
2. 海军军医大学(第二军医大学)第一附属医院病理科, 上海 200433;
3. 复旦大学附属华山医院病理科, 上海 200040;
4. 上海交通大学医学院附属新华医院病理科, 上海 200092;
5. 复旦大学附属中山医院病理科, 上海 200032
*通信作者)
摘要:
目的 评价Masson三色染色的染色缺陷,分析造成染色缺陷的主要原因和解决方案。方法 选取海军军医大学(第二军医大学)第三附属医院病理科2020年病理诊断为结节性肝硬化的石蜡标本5例。将5例石蜡标本连续切片,制成4 μm厚的切片,分发至各参评医院进行Masson三色染色。由2名主任医师按评分标准进行评分,取评分均值对切片进行定级。结果 本次Masson三色染色室间质量控制测评共发放切片50张,最终48家医院提交了染色切片(48张切片),其中甲级片17张、乙级片25张、丙级片4张、丁级片2张。出现的染色缺陷主要包括:细胞核淡染,与细胞质对比差;细胞核染色过深而模糊,细胞质呈灰蓝色;细胞质染色过深或过浅;平滑肌纤维未染色或呈灰色而非红色;组织结构呈现错误的染色或染色不均匀,如胶原蛋白未着色或整个切片染成蓝色等假象。结论 Masson三色染色操作中需严格掌握和控制染色过程中的染色温度、染色时间、pH值、试剂配制与保存等细节,从而保证三色染色的质量及染色的稳定性和可靠性。
关键词:  三色染色  组织化学  质量控制  室间质量评价
DOI:10.16781/j.CN31-2187/R.20220160
投稿时间:2022-02-24修订日期:2022-04-15
基金项目:国家自然科学基金(81472278).
Analysis of common problems and quality control of Masson trichrome staining
ZHOU Ying-ying1,XIAN Zhi-hong1*,ZHANG Shun-min2,GAO Ming-shi3,YU Wen-wei4,Su-jie AKESU5
(1. Department of Pathology, The Third Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200438, China;
2. Department of Pathology, The First Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200433, China;
3. Department of Pathology, Huashan Hospital, Fudan University, Shanghai 200040, China;
4. Department of Pathology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China;
5. Department of Pathology, Zhongshan Hospital, Fudan University, Shanghai 200032, China
*Corresponding author)
Abstract:
Objective To evaluate the staining defects of Masson trichrome staining, and analyze the main causes and solutions. Methods Five paraffin specimens of nodular liver cirrhosis diagnosed by pathology were selected in 2020 from Department of Pathology, The Third Affiliated Hospital of Naval Medical University (Second Military Medical University). They were consecutively sectioned into 4 μm slices, and then the slices were distributed to all the participating hospitals for Masson trichrome staining. The stained slices were scored by 2 physicians according to the scoring standard, and then they were graded by average score. Results In this Masson trichrome staining inter-laboratory quality control evaluation, a total of 50 slices were distributed, and finally 48 hospitals submitted stained slices (48 slices), including 17 in grade A, 25 in grade B, 4 in grade C, and 2 in grade D. The staining defects mainly included:light staining of nucleus, poor contrast with cytoplasm; the nuclei were stained too dark and fuzzy, and the cytoplasm was gray blue; the cytoplasm was stained too dark or too pale; the smooth muscle fibers was not stained or gray rather than red; and the tissue structure showed false staining or uneven staining, such as the collagen was not stained or the whole section was stained blue. Conclusion It is necessary to strictly control the staining temperature, staining time, pH value, reagent preparation and preservation and other relevant details in the staining process of Masson trichrome staining, so as to ensure the quality of trichrome staining and the stability and reliability of staining.
Key words:  trichrome staining  histochemistry  quality control  external quality assessment