斑马鱼幼鱼整鱼石蜡切片制作  

Preparation of paraffin-embedded sections of zebrafish larva

作　　者：何嘉玲 暴国 王天奇 张长勇 李楠 孙德明 HE Jialing;BAO Guo;WANG Tianqi;ZHANG Changyong;LI Nan;SUN Deming(Experimental Animal Center of National Research Institute for Family Planning,Beijing 100081,China)

机构地区：[1]国家卫生健康委科学技术研究所实验动物中心,北京100081

出　　处：《中国比较医学杂志》2020年第8期35-41,共7页Chinese Journal of Comparative Medicine

基　　金：2019年度国家卫生健康委科研所中央级公益性科研院所基本科研业务费专项(2019GJM03);2018年度国家卫计委科研所科技创新重点项目(2018GJ07)。

摘　　要：目的探讨斑马鱼幼鱼整鱼的石蜡切片制作。方法以4 d斑马鱼幼鱼为材料,探讨使用不同固定剂和脱水剂的切片效果。结果4%PFA过夜不适合幼鱼脑组织和肌肉固定,组织脆裂严重,但对肠道影响较小。Bouin’s液适合幼鱼固定,后续使用叔丁醇脱水稍优于乙醇。Bouin’s液固定的幼鱼脑组织脱水1 min,肌肉组织脱水4 min能获得良好的切片效果。结论在优化条件下,制作的切片基本达到了切片完整,薄厚均匀,无褶无刀痕,裂痕;染色核质分明,红蓝适度,透明洁净等要求。可供需进行斑马鱼幼鱼石蜡切片的研究者参考。Objective To develop a method of paraffin-embedded section preparation of whole zebrafish larva.Methods At 4 days post-fertilization,zebrafish larvae were used to explore the effect of two kinds of fixative solution and dehydration reagent on paraffin-embedded sections.Results Overnight 4%PFA fixation was not suitable for brain or muscle tissues of zebrafish larva.The tissue was severely brittle,but it had little effect on the intestines.Bouin’s fixative solution was suitable for fixation of zebrafish larva,and subsequent use of tert-butanol or ethanol showed little difference.The dehydration by tert-butanol was slightly better than that of ethanol.For Bouin’s solution-fixed zebrafish larva,brain tissue dehydrated for 1 minute and muscle tissue dehydrated for 4 minutes can obtain a standard paraffin section which has no folds,no cracks and clearly dyed.Conclusions Under optimized conditions,the prepared sections met the standard for paraffin-embedded sections:a complete,uniform thickness,no folds,no knife marks or cracks,a clearly dyed nucleus,moderately red and blue,transparent,and clean.This method can be used as a reference for researchers who need to prepare paraffin-embedded sections of zebrafish larva.

关 键 词：斑马鱼幼鱼 石蜡切片 固定剂 脱水剂 

分 类 号：R-33[医药卫生]