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目的比较环保透明脱蜡液Van-Clear和二甲苯透明脱蜡制作的组织切片苏木精-伊红(HE)染色优良率,对比荧光原位杂交(FISH)法检测2种透明脱蜡液制作的乳腺癌组织切片的人类表皮生长因子受体2(HER2)基因扩增阳性率,探讨Van-Clear替代二甲苯的可行性。方法收集中山市博爱医院2013年2月~2015年12月门诊及住院部送检的乳腺浸润性导管癌标本89例、乳腺增生标本125例、子宫平滑肌瘤标本193例、子宫平滑肌肉瘤标本12例、肺癌穿刺标本16例、绒毛标本139例,同一病变部位切取2个样本,用抽签法随机分为2组,命名为A、B组。A组采用二甲苯透明脱蜡制作切片574张;B采用Van-Clear透明脱蜡制作切片574张。依据切片染色情况判定切片等级,采用SPSS20.0软件比较A、B组切片HE染色优良率;A、B组中乳腺浸润性导管癌标本再各制作切片89张,采用SPSS 20.0软件比较FISH法检测的HER2基因扩增的差异。结果 (1)生物显微镜下,A、B两组切片HE染色结果细胞轮廓清晰、透明度好,细胞核与细胞质蓝红相映、色彩鲜艳,核质对比明显,核膜及核染色质颗粒清晰可见、分辨良好,分裂象染色体呈现黑蓝色,各种成分层次分明。(2)A、B两组切片HE染色优良片分别为570张、572张,中等、差片4张、2张。染色优良率分别为99.30%、99.65%,两组间染色优良率差异无统计学意义(χ2=0.50,P>0.05)。(3)荧光显微镜下,A、B两组切片乳腺浸润性导管癌HER2双探针FISH检测结果组织轮廓和背景均清晰,探针定位准确,可见耀眼的红/绿荧光信号。(4)A、B两组切片检测乳腺浸润性导管癌HER2基因,FISH阳性率分别为24.72%、26.97%,两组间阳性率差异无统计学意义(χ2=0.50,P>0.05),且FISH结果符合率为97.75%。结论环保透明脱蜡液Van-Clear有替代传统试剂二甲苯应用于组织切片HE染色及FISH法检测乳腺癌HER2基因的潜在可能。
Objective To compare the excellent rate of hematoxylin-eosin (HE) staining on the tissue slices made by the environmentally-friendly transparent dewaxing liquid Van-Clear and xylene transparent dewax, and to compare the production of two transparent dewaxing liquids by fluorescence in situ hybridization (FISH). The positive rate of human epidermal growth factor receptor 2 (HER2) gene amplification in breast cancer tissue sections was investigated to investigate the feasibility of Van-Clear substitution of xylene. Methods A total of 89 invasive ductal carcinoma specimens, 125 breast hyperplasia specimens, 193 uterine leiomyoma specimens, and 12 uterine leiomyosarcoma specimens were collected from the outpatient and inpatient departments of the Bo’ai Hospital, Zhongshan City from February 2013 to December 2015. Cases, 16 cases of lung cancer puncture specimens, 139 cases of villous specimens, the same lesions were taken 2 samples, were randomly divided into 2 groups by drawing method, named A, B group. In group A, 574 slices were made using xylene transparent dewaxing, and 574 slices were made using Van-Clear transparent dewaxing. According to the section staining, the grade of the slice was determined. The excellent rate of HE staining was compared between group A and group B using SPSS20.0 software. The invasive ductal carcinoma specimens of group A and group B were again prepared with 89 slices. The results were compared by FISH method using SPSS 20.0 software. The difference in HER2 gene amplification. Results (1) Under the biomicroscope, the HE staining results of the two groups A and B showed clear cell outlines and good transparency. The nucleus and cytoplasm were blue and red, and the color was bright. The nucleocytoplasm was obviously contrasted. Nuclear membranes and nuclear chromatin particles were clearly visible and resolved. Good, split chromosomes appear black and blue, and various components are structured. (2) A, B two groups of slices of HE staining were 570, 572, medium, poor, 4 and 2 sheets. The excellent and good staining rates were 99.30% and 99.65%, respectively. There was no significant difference in the excellent rate of staining between the two groups (χ2=0.50, P>0.05). (3) Under the fluorescence microscope, the HER2 double-probe FISH detection results of the two groups of A and B sections were clear, the tissue outline and background were clear, the probe positioning was accurate, and dazzling red/green fluorescence signals were visible. (4) The HER2 gene of breast infiltrating ductal carcinoma was detected in both groups A and B. The positive rate of FISH was 24.72% and 26.97% respectively. There was no significant difference between the two groups (χ2=0.50, P>0.05). The coincidence rate of FISH results was 97.75%. Conclusion Van-Clear, an environmentally friendly transparent dewaxing liquid, has the potential to replace the traditional reagent xylene in HE slice staining and FISH detection of HER2 gene in breast cancer.