【作者】 赵颖； 赵德明； 王剑伟； 李爱华； 杨利峰；

【Author】 Zhao Ying;Yang Lifeng;Zhao Deming;College of Vererinary Medicine,China Agriculture University;Instiute of Hydrobiology,The Chinese Academy of Science;

【机构】 中国农业大学动物医学院； 中国科学院水生生物研究所；

【摘要】 目的目前为止鲜有专门针对稀有鮈鲫这种新型实验动物进行组织切片制作方法的研究,故我们对此进行了实践摸索以建立一完整的组织学切片制作流程供参考交流。方法对正常生活状态下的健康稀有鮈鲫采用Bouin’s固定液固定,并用脱钙液软化骨骼后进行修块,组织块经梯度酒精脱水、松油醇透明之后,按照标准流程制成石蜡切片,HE染色后封片用显微镜观察。结果:切片主要采取横断面和矢状面作为观察方位,并使用显微拍摄装备摄取了各系统组织脏器的组织学照片,视野良好结构分明。结论本文所述的方法经过多次探索和实践,制得的组织切片镜检结果清晰,质量稳定,可为以稀有鮈鲫等小型硬骨鱼为实验动物的切片制作提供一些可行的建议。更多还原

【Abstract】 Background Gobiocypris rams,a freshwater fish with high prolificacy,distributes in Sichuan Province.In order to be a laboratory animal,the fish has been investigated its geographical distribution,morphology,habita,ontogenesis,reproduction,growth,taxonomy,semitiviqto ecological factors,karyotype and so on by Chinese Academy of Science,since 1990 s.The length of the fish is 38 mm to 85 mm.The fish,has high adaptability to the temperature,oxygen and CO2 in fresh water,has been used in many experiments of environmental science research and genetics research.However,there were few histological background of Gobiocypris rams until now.Here,we introduce a modified way to make sections for a better view.Methods The healthy individuals were fixed in the Bouin’s fluid for 24-48 hrs.Then,wash the fish with running water slightly,and put the samples into decalcifying fluid.6-16 hrs later,after finishing decalcification,the fish were cut into several small pieces(preferably inferior to 0.3-0.8 cm3),the specimen was dehydrated with 50%,75%,85%,95%alcohol and terpilenol in order,and then waxed with hot melt paraffin.When infiltration was completed,cold the specimen below freezing temperature.The specimen in solid paraffin was allowed to cut with knife into serial slice(3-5μm).The slices were dyed by Hematoxylin and Eosin(H&E) staining according to the routine protocol.The sections are then placed onto glass microscope slides to protect the tissue and provide a better view.Transverse and sagittal section pictures of the fish were collected by a optical microscope.Results The sections were scaned under microscope and the pictures of different kinds of organs and tissues were taken.Those high quality picture of histology can fill the background of Gobiocypris rams.Conclusion Histology can detect features of disease which are difficult to recognize from clinical symptoms.The normal histology of Gobiocypris rams tissues and organs is the foundation of pathology knowledge base.The way to make a better section of Gobiocypris rams may contribute to the research of aquatic animal.更多还原