人脑胶质瘤组织双色荧光染色图像质量与诊断效能分析

Quality and diagnostic performance of images of dual-color fluorescence staining of human glioma

目的分析人脑胶质瘤组织双色荧光染色的图像质量与诊断效能,并与HE染色进行比较。方法取新鲜人脑胶质瘤组织标本71例,将同一脑组织标本分别分成大小相同的两份。一份标本使用荧光染色剂BODIPY和DRAQ5分别染脑组织脂质和DNA,在激光扫描共聚焦显微镜下进行双色荧光成像,采集荧光图像。另一份标本进行HE组织染色,用玻片扫描仪进行扫描成像。分别于染色后及染色90 d后由两名病理科医生对双色荧光染色和HE染色组织图像进行评估。对两位病理医生的评估结果进行Cohen'skappa一致性检验,评价双色荧光染色与HE染色的诊断一致性,计算双色荧光染色的诊断准确率、敏感度、特异度。结果人脑胶质瘤组织双色荧光染色图像具有与HE染色图像相似的组织学特征,且脂质保留完整。病理科医生A评估双色荧光染色图像得到低级别胶质瘤21例、高级别胶质瘤50例,病理科医生B分别为24、47例,将病理科医生A、B第一次诊断结果与HE染色诊断结果进行Kappa一致性检验,κ值分别为0.48、0.64。染色90 d后,两位病理科医生第二次评估双色荧光染色图像,病理科医生A评估低级别胶质瘤26例、高级别胶质瘤45例,病理科医生B分别为28、43例,将病理科医生A、B第二次诊断结果与HE染色诊断结果进行Kappa一致性检验,κ值分别为0.76、0.82。病理科医生A第一次诊断敏感度为88%、特异度为57%、诊断准确率为76%,第二次诊断分别为93%、82%、89%;病理科医生B第一次诊断敏感度为91%、特异度为71%、诊断准确率为83%,第二次诊断分别为93%、89%、92%。结论人脑胶质瘤组织的双色荧光染色图像具有类似HE染色的组织学特征,且能够完整保留脂质;同时与HE染色相比,诊断一致性基本满意,诊断准确率、敏感度及特异度高。

Objective To analyze the quality and diagnostic performance of images of dual-color fluorescence stain-ing of human glioma and to compare dual-color fluorescence staining with HE staining.Methods Seventy-one cases of fresh human brain tissue specimens were collected and divided into two samples of the same size for dual-color fluorescence staining and HE staining.The lipids and DNA of brain tissues in one sample were stained with BODIPY and DRAQ5,re-spectively,and fluorescence imaging was performed under laser scanning confocal microscope,and we collected the fluo-rescence images.The fluorescence images were randomly evaluated by two pathologists and were compared with the images of HE staining.Another sample was stained with HE and scanned with a slide scanner.Two pathologists evaluated the im-ages of dual-color fluorescence staining and HE staining separately after staining and evaluated again after 90 days of dual-color fluorescence staining.The consistency test was performed on the result of evaluation of the two pathologists andκval-ue was calculated to judge the diagnostic consistency of dual-color fluorescence staining and HE staining.The diagnostic accuracy rate,sensitivity,and specificity of dual-color fluorescence staining were calculated.Results The fluorescence images of dual-color fluorescence staining of human glioma showed similar histological characteristics to images of HE staining,and lipids of dual-color fluorescence staining were remained complete.Pathologist A obtained 21 cases of low-grade glioma and 50 cases of high-grade glioma by evaluating the images of dual-color fluorescence staining.Pathologist B obtained 24 cases and 47 cases,respectively.Kappa consistency test was performed between the first diagnosis results of pathologists A and B and the diagnosis results of HE staining,andκvalue was 0.48 and 0.64,respectively.After 90 days of dual-color fluorescence staining,two pathologists evaluated the images of dual-color fluorescence staining for the second time.In this time,pathologist A obtained 26 cases of low-grade glioma and 45 cases of high-grade glioma and pathologist B evaluated 28 and 43 cases,respectively.Kappa consistency test was performed by comparing the second diagnosis result of pathologists A and B with the diagnosis result of HE staining,andκvalues were 0.76 and 0.82,respectively.The sen-sitivity,specificity,and diagnostic accuracy of pathologist A were 88%,57%and 76%in the first diagnosis,and 93%,82%and,89%in the second diagnosis,respectively.The sensitivity,specificity and accuracy of pathologist B were 91%,71%,and 83%in the first diagnosis,and 93%,89%and 92%in the second diagnosis,respectively.Conclusions The images of dual-color fluorescence staining of human glioma have similar histological characteristics to images of HE stain-ing,and dual-color fluorescence staining remains lipids complete.Compared with HE staining,the diagnostic consistency of dual-color fluorescence staining is basically satisfied and it has high diagnostic accuracy,sensitivity,and specificity.