8种染色方法对组织切片中虾肝肠胞虫染色效果的比较

病理检测是一种诊断病原及观察机体组织病理变化的重要方法,特殊染色是在病变组织中寻找病原体的前提手段,良好的染色有助于将病原体与组织区别,从而增加病原在组织切片中的检出率。为筛选出一种适用于虾肝肠胞虫(Enterocytozoon hepatopenaei,EHP)病理切片检测的最佳染色方法,使用苏木精-伊红(HE)染色法、Masson染色法、爱显蓝-雪夫(AB-PAS)染色法、甲苯胺蓝(TB)染色法、伟郝夫范吉森(EVG)染色法、劳克坚牢蓝(LFB)染色法、天狼猩红(Sirius red)染色和普鲁士蓝(PB)等8种染色方法对凡纳滨对虾(Litopenaeus vannamei)感染了虾肝肠胞虫的肝胰腺进行组织病理切片染色,并结合对EHP检出率的统计分析,比较了这些染色方法的优缺点。结果表明:HE染色法、AB-PAS染色法、TB染色法、EVG染色法、天狼猩红Sirius red染色法和PB染色法对孢子的染色效果较差,与背景颜色对比不明显,单个视野检出率较低; LFB染色法虽然使孢子易被识别,但是有孢子丢失现象,且组织染色效果差; Masson染色结果显示孢子呈现品红色,与组织颜色对比鲜明,孢子颗粒清晰可见,对组织细胞结构染色效果较好,且检出率也最高。综合分析染色效果及检出率,Masson染色法是一种非常适用于组织内的虾肝肠胞虫孢子染色的方法,本研究为微孢子虫病的组织病理研究提供一种有效的基础手段。

Comparison of 8 different special staining techniques of Enterocytozoon hepatopenaei in histologic section

Histopathological examination is an important method for diagnosing pathogens and observing histopathological changes of the organism.Special staining is the prerequisite for finding pathogens in infected tissues.Good staining resluts can help to distinguish pathogens from tissues,and increase the detection rate of pathogens in tissue sections.In order to screen out a suitable staining method for the detection of Enterocytozoon hepatopenaei(EHP)in histopathology,8 different staining methods,including:Hematoxylin-eosin staining,Masson staining,Alcian blue-Periodic acid schiff staining,Toluidine blue staining,Verhoeff-Van Gieson staining,Luxol fast blue staining,Sirius red and Prussian blue staining method,were selected to compare the dyeing effects using the EHP infected hepatopancereas tissues of Penaeus vannamei.Combined with the detection rate of EHP,the advantages and disadvantages of these staining methods were compared.The dyeing effects of HE,AB-PAS,TB,EVG,Sirius red and PB methods were poor,the contrast between the spores and the background was not noticeable,the detection rates of a single visual field were low.Although LFB method made the spores easy to be identified,it had spores loss and bad staining effect on host tissues.Masson method showed that the spores color was clearly visible magenta,the staining effects of the tissue cell structure were better,and the detection rate of the single field was the highest in the 8 methods.Comprehensive analysis of the effects of staining and detection rates showed that Masson method was very well suitable for the staining of EHP spores in host tissues.This study provides an effective basic method for the histopathological study of microsporidiosis.