斑马鱼幼鱼整鱼石蜡切片制作
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国家卫生健康委科学技术研究所实验动物中心,北京 100081

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Preparation of paraffin-embedded sections of zebrafish larva
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Experimental Animal Center of National Research Institute for Family Planning, Beijing 100081, China

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    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。

    Abstract:

    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.

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何嘉玲,暴 国,王天奇,张长勇,李 楠,孙德明.斑马鱼幼鱼整鱼石蜡切片制作[J].中国比较医学杂志,2020,30(8):35~41.

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  • 收稿日期:2020-06-18
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  • 在线发布日期: 2020-09-02
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