日本血吸虫尾蚴不同染色方法的比较研究

Comparative study on different staining methods of Schistosoma japonicum cercariae

目的探索不同化学染色法对日本血吸虫尾蚴的染色效果,并对其形态结构进行分析,为血吸虫研究和教学提供参考。方法 采用自然逸出法从阳性钉螺获取尾蚴,涂片法制作尾蚴玻片标本,分别采用苏木素-伊红(HE)染液、瑞氏-姬姆萨染液、尼氏(Nissl)染液、0.4%台盼蓝、考马斯亮蓝R-250染液、丽春红染液、0.1%茜素红染液、刚果红染液等化学染料进行染色,在光学显微镜下观察和评价尾蚴形态结构,比较各方法的染色效果。结果形态学测量的结果显示获取的尾蚴为成熟期尾蚴,8种染色方法均能使血吸虫尾蚴呈不同程度的着色,但在尾蚴形态结构显示上有相似性,亦有不同之处。HE染色、瑞氏-姬姆萨染色、台盼蓝和尼氏染色等对尾蚴头部和尾部结构均能较好显示。考马斯亮蓝和丽春红染色对尾蚴头部微细结构均显示欠佳,但考马斯亮蓝可显示尾部排泄管,丽春红可显示尾部部分结构。茜素红、刚果红染色效果类似,对头部和尾部结构均能部分显示。结论 HE染色能将尾蚴各部位器官显示清楚,对比明显,可在教学科研中广泛使用。其余染色方法应根据研究目的不同,选取不同的方法。

Objective Explore the dyeing effect of different chemical staining methods on cercariae of Schistosoma japonicum and to identify the morphological structure of cercariae,so as to provide reference for the research and teaching of Schistosoma japonicum. Methods Cercariae were obtained from positive oncomelania by natural escape way,and cereariae slides were prepared by smear method.Chemical staining was performed with hematoxylin-eosin(HE),Reiss-Giemsa,Nissl,0.4% trypan blue,coomassie brilliant blue R-250,Lichun red,0.1% alizarin red,Congo red,etc.The morphological structure of cercariae was observed and evaluated under lightmicroscope,comparing the effect of dyeing method. Results Morphological measurements showed that the cercariae collected were mature cercariae.All the eight staining methods could stain the cercariae of Schistosoma japonicum to varying degrees,but there were similarities and differences in the morphological structure of cercariae.HE staining,Wright-Giemsa staining,Trypan blue staining and Nissl staining all showed the head and tail structure of cercariae well;Coomassie brilliant blue staining and Lichun red staining did not show structure of cercariae head,but Coomassie brilliant blue could show the tail excretory tube,Lichun red could show the tail structure.Alizarin red and Congo red staining have similar effects,and can partially display the structure of head and tail. Conclusion HE staining can show better the organs of cercariae clearly,and can be widely used in teaching and researching.Other dyeing methods should be chosen according to different research purposes.