实验动物与比较医学 ›› 2024, Vol. 44 ›› Issue (6): 675-681.DOI: 10.12300/j.issn.1674-5817.2024.049

• 动物实验技术与方法 • 上一篇    下一篇

6种复合固定液对金黄仓鼠视网膜组织固定效果的比较

吴海凤(), 周小江, 李晨江, 李怀银, 高明()()   

  1. 兰州生物制品研究所有限责任公司, 甘肃省疫苗工程技术研究中心, 兰州 730046
  • 收稿日期:2024-03-28 修回日期:2024-07-03 出版日期:2025-01-04 发布日期:2024-12-25
  • 通讯作者: 高 明(1992—),男,助理研究员,研究方向:实验动物学。E-mail: gaominglss@163.com。ORCID: https://orcid.org/0009-0003-2618-6587
  • 作者简介:吴海凤(1977—),女,硕士,医学生物工程师,研究方向:实验动物病理检测。E-mail: 1092999471@qq.com

Comparison of the Fixation Effects of Six Composite Fixatives on Retinal Tissue of Golden Hamsters

WU Haifeng(), ZHOU Xiaojiang, LI Chenjiang, LI Huaiyin, GAO Ming()()   

  1. Lanzhou Institute of Biological Products Co. , Ltd. , Gansu Vaccine Engineering Technology Research Center, Lanzhou 730046, China
  • Received:2024-03-28 Revised:2024-07-03 Published:2024-12-25 Online:2025-01-04
  • Contact: GAO Ming (ORCID: 0009-0003-2618-6587), E-mail: gaominglss@163.com

摘要:

目的 比较6种复合固定液对金黄仓鼠视网膜组织石蜡切片的影响,优化金黄仓鼠视网膜组织石蜡切片的固定方法。 方法 取18只雄性SPF级金黄仓鼠,随机分为6组,每组3只。每只动物经腹腔注射戊巴比妥钠麻醉后,分别使用4%多聚甲醛、Bouin's、Carnoy、Davidson's、Zenker和Helly固定液进行心脏灌注固定。将每只动物的眼球组织取出后制作眼杯,并投入对应的复合固定液中固定,48 h后取出进行石蜡包埋,并将包埋块在切片机下切片,然后进行苏木精-伊红(hematoxylin and eosin,HE)染色。在光学显微镜下观察视网膜组织细胞的形态学特征并进行双盲法评分,统计分析各种复合固定液的固定效果和染色质量。 结果 Davidson's固定液固定后的切片中,视网膜完整无断裂,各层结构清晰,细胞核形态良好,视网膜与巩膜、葡萄膜连接紧密;4%多聚甲醛、Bouin's和Carnoy固定液固定的切片中均有不同程度的视网膜断裂以及视网膜与巩膜脱离现象;Zenker和Helly固定液固定后的切片质量最差,视网膜组织脱落明显,裂隙大,各层细胞不清晰,染色淡。对视网膜细胞形态特征进行双盲法评分后,SPSS 27.0软件分析结果显示:6种固定液的平均得分差异有统计学意义(P<0.001),其中Davidson's固定液>4%多聚甲醛>Bouin's固定液>Carnoy固定液>Helly固定液>Zenker固定液;Zenker固定液和Helly固定液与其他4种试剂的平均得分相比均有显著差异(P<0.001),其他试剂之间两两相比无明显差异(P>0.05)。 结论 Davidson's固定液染色效果最佳,4%多聚甲醛和Bouin's固定液染色效果次之,Carnoy固定液染色效果较差,Zenker固定液和Helly固定液染色效果最差。据此提示,金黄仓鼠视网膜组织的固定首选Davidson's固定液,其次为4%多聚甲醛、Bouin's和Carnoy固定液。

关键词: 固定液, 金黄仓鼠, 视网膜, 石蜡切片

Abstract:

Objective To compare the effects of six composite fixatives on paraffin sections of golden hamster retinal tissue, and to optimize the fixation methods for retinal tissue paraffin sections of golden hamsters. Methods Eighteen male SPF grade golden hamsters were taken and randomly divided into six groups of three animals each. After each animal was anesthetized by intraperitoneal injection of sodium pentobarbital, cardiac perfusion was performed using 4% paraformaldehyde, Bouin's, Carnoy, Davidson's, Zenker, and Helly fixatives, respectively. The eye tissues of each animal were taken out to make eye cups and put into the corresponding compound fixative solution for fixation, and then taken out for paraffin embedding after 48 h. The embedded blocks were sliced using microtome, and then stained with hematoxylin and eosin (HE). The morphological characteristics of retinal tissue cells were observed under a light microscope and scored in a double-blind method to statistically analyze the fixation effect and staining quality of various composite fixative solutions. Results Retinal sections fixed with Davidson's fixative exhibited intact morphology without breaks, clear structural layers, well-morphosed nuclei, and tight adhesion between the retina, sclera, and uvea. In contrast, sections fixed with 4% paraformaldehyde, Bouin's, and Carnoy fixatives showed varying degrees of retinal breaks and detachment from the sclera. Sections fixed with Zenker and Helly fixatives demonstrated the poorest quality, characterized by pronounced detachment, large fissures, unclear cell layers, and pale staining. Statistical analysis using SPSS 27.0 software revealed significant differences in mean scores among the six fixatives (P<0.001). The fixation quality ranking was as follows: Davidson's fixative > 4% paraformaldehyde > Bouin's fixative > Carnoy fixative > Helly fixative > Zenker fixative. Zenker and Helly fixatives showed significantly lower scores than the other fixatives (P<0.001), while no significant differences were observed among the remaining fixatives (P>0.05). Conclusion Davidson's fixative provides the best fixation and staining results, followed by 4% paraformaldehyde and Bouin's fixative. Carnoy fixative exhibits suboptimal performance, while Zenker and Helly fixatives result in the poorest outcomes. Therefore, Davidson's fixative is recommended as the optimal fixative for golden hamster retinal tissue, with 4% paraformaldehyde, Bouin's and Carnoy fixatives as alternative options.

Key words: Fixative solution, Golden hamster, Retinal, Paraffin section

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