探讨适用于HE染色、免疫组织化学染色和分子检测的最佳脱色素方法  

Optimal melanin removal methods for HE staining,immunohistochemistry and molecular detection

作　　者：张文雯 邱雅婷 吴晨榆 柯龙凤 朱伟峰[1] 陈刚[1] 陈燕坪[1] Zhang Wenwen;Qiu Yating;Wu Chenyu;Ke Longfeng;Zhu Weifeng;Chen Gang;Chen Yanping(Department of Pathology,Clinical Oncology School of Fujian Medical University,Fujian Cancer Hospital,Fuzhou 350014,China;Department of Molecular Pathology,Clinical Oncology School of Fujian Medical University,Fujian Cancer Hospital,Fuzhou 350014,China)

机构地区：[1]福建医科大学肿瘤临床医学院福建省肿瘤医院病理科,福州350014 [2]福建医科大学肿瘤临床医学院福建省肿瘤医院分子病理研究室,福州350014

出　　处：《中华病理学杂志》2024年第6期570-577,共8页Chinese Journal of Pathology

基　　金：福建医科大学启航基金(2021QH1140)。

摘　　要：目的探索适用于HE染色、免疫组织化学染色和分子检测的最佳脱色素方法。方法收集2018年1月至2022年3月福建省肿瘤医院病理科38例富含黑色素的黑色素瘤石蜡标本,制成组织芯片,分别采用水浴过氧化氢法、双重氧化脱色素法、改良高锰酸钾法、三氯异氰尿酸法脱黑色素后行HE染色。之后将切片分成2个队列,第1个队列分别采用上述4种方法脱色素后再行免疫组织化学染色(SOX10、Ki-67、HMB45和Melan A);第2个队列先行同样指标的免疫组织化学染色后再分别采用上述4种方法脱色素。另收集其中色素含量较高的17例石蜡标本,采用上述4种方法脱黑色素后提取DNA,进行DNA浓度及质量的测定。比较分析不同脱色素方法的脱色素能力,以及对HE和免疫组织化学染色的效果、DNA浓度和质量的影响。结果在脱黑色素能力方面,改良高锰酸钾法和水浴过氧化氢法脱色素效果最好,均仅5.26%(2/38)有少量色素残留;其次是三氯异氰尿酸法,10.53%(4/38)有大量色素残留;最差的是双重氧化脱色素法,15.79%(6/38)有大量色素残留。在HE染色方面,水浴过氧化氢法脱色素后染色好的比例达92.11%,优于其他3种方法。在免疫组织化学染色方面,改良高锰酸钾法、双重氧化脱色素法和三氯异氰尿酸法,先免疫组织化学染色后脱色素的染色评分均值分别为20.84、26.63和35.02,均优于先脱色素后免疫组织化学的染色评分均值(8.70、15.41和21.22);水浴过氧化氢法,先脱色素后免疫组织化学的染色评分均值为33.57,优于先免疫组织化学染后脱色素的染色评分均值(19.96)。HMB45、Melan A和Ki-67这3个指标采用先免疫组织化学染色后三氯异氰尿酸脱色素的染色评分分别为36.45、33.79和36.24,效果最佳;SOX10采用先水浴过氧化氢法脱色素后免疫组织化学染色的评分为34.39,效果最佳。在DNA浓度及质量方面,采用双重氧化脱色素法、改良高�Objective To seek the optimal melanin-removal method for hematoxylin and eosin(HE)staining,immunohistochemistry and molecular detection.Methods Thirty-eight paraffin tissue samples of malignant melanoma diagnosed at the Fujian Cancer Hospital,Fuzhou,China between January 2018 and March 2022 were collected and used to make a tissue microarray.Melanin in these cases was removed using warm hydrogen peroxide,double oxidation depigmentation,modified potassium permanganate-oxalic acid or trichloroisocyanuric acid,followed by HE staining.The cases were divided into two cohorts:one was subject to the one of the above four methods to remove melanin first,followed by immunohistochemistry(SOX-10,Ki-67,HMB45 and Melan A),while the other was subject to immunohistochemical staining first and then a melanin removal.Following that,seventeen melanin-rich paraffin tissue samples were collected and depigmented using the methods described above.DNA extraction was then done,followed by assessments of DNA content and quality.Moreover,the completeness of melanin removal,the effect on HE and immunohistochemical staining,and the quality of DNA were compared between the depigmented methods.Results Regarding the effectiveness of melanin removal,the modified potassium permanganate-oxalic acid and the warm hydrogen peroxide methods were the most effective,and both showed residual melanin in only 5.26%(2/38)of the cases.The trichloroisocyanuric acid method showed residual melanin in 10.53%(4/38)of the cases.The worst was the double oxidation depigmentation method,which showed pigment residue in 15.79%(6/38)of the cases.For HE staining,the percentage of good staining with the warm hydrogen peroxide method was 92.11%,higher than the other three methods.For immunohistochemical staining,the mean staining scores of immunohistochemistry first followed by melanin removal with modified potassium permanganate-oxalic acid,double oxidation and trichloroisocyanuric acid were 20.84,26.63 and 35.02,respectively.These immunohistochemical staining scores wer

关 键 词：黑色素瘤 分子诊断技术 免疫组织化学 

分 类 号：R361.2[医药卫生—病理学]