深圳大学学报理工版

许特,王筠,邓芳,等.组织芯片多重免疫组织化学染色方法的优化[J].深圳大学学报理工版,2019,36(No.5(473-598)):570-575.[doi:10.3724/SP.J.1249.2019.05570]
XU Te,WANG Yun,DENG Fang,et al.Technique improvement of multiple immunohistochemical staining on tissue microarray[J].Journal of Shenzhen University Science and Engineering,2019,36(No.5(473-598)):570-575.[doi:10.3724/SP.J.1249.2019.05570]

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组织芯片多重免疫组织化学染色方法的优化

许特^1,2,3,王筠¹,邓芳^1,4,李延鹏¹,屈军乐²,颜鸿飞⁴,盛司潼¹

1)深圳大学生命与海洋科学学院,广东深圳518055; 2)深圳大学物理与光电工程学院,光电子器件与系统教育部/ 广东省重点实验室,广东深圳518060; 3)西安交通大学口腔医院,陕西省颅颌面精准医学重点实验室, 陕西西安710004; 4)汕头大学医学院附属肿瘤医院,广东汕头 515000

关键词：分子病理学; 组织芯片; 免疫组织化学法; 多重染色技术; 抗原修复

Technique improvement of multiple immunohistochemical staining on tissue microarray

XU Te^{1, 2, 3}, WANG Yun¹, DENG Fang^{1, 4}, LI Yanpeng¹, QU Junle², YAN Hongfei⁴, and SHENG Sitong¹

1)College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, Guangdong Province, P.R.China2)College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China3)College of Stomatology, Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, Xi'an Jiaotong University, Xi'an 710004, Shaanxi Province, P.R.China4)Cancer Hospital, Shantou University Medical School, Shantou 515000, Guangdong Province, P.R.China

Keywords： molecular pathology; tissue microarray; immunohistochemistry; multiple staining; antigen retrieval

DOI: 10.3724/SP.J.1249.2019.05570

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为提高多重免疫组织化学(immunohistochemistry, IHC)技术在组织芯片(tissue microarray, TMA)上的染色效果,通过对同一样本进行多种抗体染色或同种抗体多次染色的方法,探索并优化IHC技术中的多个步骤,具体包括:“滤纸夹层-蒸蛋器”抗原修复新装置的研发、抗原修复液的筛选、透明剂的优选. 研究结果显示:自创的“滤纸夹层-蒸蛋器”抗原修复辅助工具,能减少高温高压修复时水分子对组织的冲击,作用温和、操作安全,有较好的组织切片完整性和较少的组织脱片率,且可减少抗原修复液使用量; 柠康酐抗原修复液对多重染色具有更强的阳性效应和通用性; 无毒TO(terpentine oil)透明剂可取代有毒二甲苯,具有同样的脱蜡透明效果.TMA中多重IHC技术的改良,提供了更加高效、廉价、安全的方法,可提升与扩展TMA在病理学中的应用价值.

To improve the technique of multiple immunohistochemistry(IHC)on tissue microarray(TMA), several steps of multiple IHC are optimized via utilizing multi-antibody or single antibody multi-staining in one microarray. The optimized methods include “filter paper interlayer-egg steamer” antigen retrieval tool, antigen retrieval buffer and transparent reagent. The results show that, using invented set of “filter paper interlayer-egg steamer”, it can mitigate the impact force of water flow during antigen retrieval process to protect tissue damage, and save antigen retrieval buffer. As antigen retrieval solution, citraconic anhydride has stronger positive effect on multiple staining and for most antibodies. As transparent solution, non-toxic terpentine oil(TO)has the same results of deparaffin and transparent effects. In conclusion, the improve of multiple IHC provides a more efficient, cheap and safe method, which can greatly enhance the value of TMA application in clinical pathology.