Metafer 5玻片自动扫描系统在循环肿瘤细胞检测中辅助诊断的应用

Application of Metafer 5 Slide Automatic Scanning System in Auxiliary Diagnosis of Circulating Tumor Cells

本研究采用新建立的循环肿瘤细胞(CTCs)精准分析模块(Metafer 5)检测临床血液样本中的CTCs。为使Metafer 5能开展精准分析,针对该数字模块进行了优化。本研究纳入34例病理确诊的恶性实体肿瘤患者,通过对比4',6-二脒基-2-苯基吲哚(DAPI)、Rhodamie、异硫氰酸荧光素(FITC)、TexRed四种荧光染色剂的特点筛选区域化最佳对焦荧光染料,并对比10×PA、10×PN和20×PA物镜对CTCs判定的准确性和扫描效率,优化各个荧光通道的曝光时间,选出Metafer 5 CTCs自动扫描模式下的最佳应用参数。另外,分别采用人工镜检和Metafer 5玻片自动扫描系统两种方式对CTCs进行判定。结果表明,DAPI为最适用于区域化自动对焦的荧光染料。两种物镜镜头扫描CTCs的时间分别为(114.70±22.08)及(116.50±47.38)min,10×PA物镜具有更高的CTCs识别准确度和效率。当将DAPI、EpCAM、CD45和CEP8曝光时间分别调整为0.003、0.800、0.040和0.068 s时,荧光图像最清晰,质量最佳。同时,经过最优化参数调节Metafer 5 CTCs精准扫描模块的系统较人工镜检时间缩短了73.40 min(P<0.05),较人工镜检效率有显著提高,且该模块总CTCs检出率可达100.00%,比人工镜检高了8.83%。经参数优化后的Metafer 5玻片自动扫描系统能自动、精准、快速判读血液中的CTCs,可用于CTCs辅助判读,具有很好的临床应用前景。

The newly established accurate analysis module for detecting circulating tumor cells(CTCs)(Metafer 5)was used in this study.In order to achieve accurate analysis by Metafer 5,the digital module was optimized.Thirty-three malignant tumors patients were enrolled.Meanwhile,in order to acquire the best application parameters by Metafer 5,we screened the best fluorescent dyes by comparing the characteristics of 4',6-diamidino-2-phenylindole(DAPI),Rhodamine,fluorescein isothiocyanate(FITC)and TexRed,assessed accuracy and efficiency of CTCs scanning among 10×PA,10×PN,and 20×PA objective lens and optimized the exposure time of each fluorescent channel.Additionally,CTCs identification was evaluated by both manual examination and Metafer 5 system.According to the data,the results showed that scanning time of CTCs by two objective lens was(114.70±22.08)and(116.50±47.38)min,respectively,and the 10×PA objective lens presented higher accuracy and efficiency.When the exposure time of DAPI,EpCAM,CD45 and CEP8 was adjusted to 0.003,0.800,0.040 and 0.068 s,respectively,the clearest and best quality fluorescent images were presented.At the same time,comparing to the manual work,based on the optimal parameters of Metafer 5,not only the time cost was 73.40 min less(P<0.05),but also the accuracy reached 100.00%,which was 8.83%higher than manual work.Above all,the optimized Metafer 5 system could detect CTCs automatically,accurately and quickly in blood sample,which could be used for auxiliary interpretation of CTCs,and could have a future application in clinical practice.