不同固定液对大鼠睾丸固定效果影响-中国试验动物学报.DOCVIP

研究报告 大鼠睾丸组织固定法的优选研究 李 雷，严建燕，苏 欣，邵 悦，程 熠，周 莉，刘向云，孙祖越 （上海市计划生育科学研究所药理毒理学研究室，中国生育调节药物毒理学检测中心，上海 200032） 【摘要】目的 选择一种最优势、合理的固定方式以提高对睾丸组织制片效果，以配合不同种类的科学研究。方法 选用10%甲醛溶液、NBF-Bouins、Bouins和改良Davidsons四种不同的固定液对大鼠睾丸进行充分固定后，制作石蜡切片，进行HE染色，比较不同固定液中的睾丸组织学形态的差异；利用糖原特殊染色（PAS），探讨不同固定液对睾丸糖原观察的影响；采用免疫组织化染色，测评睾丸组织内雄激素受体的固定效果。结果 改良Davidsons固定液较NBF-Bouins引起的曲细精管萎缩轻，形态更为清晰，用免疫组织化学方法检测雄激素更为敏感，并且改良Davidsons 固定液在需要对精子发生进行分期时，其PAS 染色的效果与Bouins液固定后等同。结论 与其它固定液相比，改良Davidoson液对大鼠睾丸的固定效果较好。 【关键词】固定液；大鼠；睾丸；Davidsons；免疫组化染色； 【中图分类号】R33【文献标识码】A【文章编号】1671-7856（201）-0000-00 Doi:10.3969/j.issn.1671-7856.2013.006.000 Selection of optimal fixative for preservation of rat testis LI Lei，YAN Jan-yan，SU Xin，SHAO Yue，CHENG Yi，ZHOU Li，LIU Xiang-yun， SUN Zu-yue (Department of Pharmacology and Toxicology, Shanghai Institute of Planned Parenthood Research，National valuation Center for the Toxicology of Fertility Regulation Drugs, Shanghai 200032) 【Abstract】 Objective Histopathological examination of testes is important in assessing the reproductive damages in drug toxicological studies. Superior fixative is an important factor for preservation of stable and clear histological features. Methods Twenty healthy male Sprague-Dawley rats in the age of 60-90 days were used in this study. The rat testes were fixed in 10% buffered formalin (NBF), 10% NBF and Bouins fluid, Bouins fluid and modified Davidsons fluid (mDF), and the morphology of the testis was compared using hematoxylin and eosin (HE)-staining, periodic acid-Schiff (PAS) staining and immunohistochemical (IHC) staining. Results Fixation in mDF resulted in more sharply defined nuclear details and better preservation of the cell cytoplasm, noticeably less shrinkage of the seminiferous tubules, and superior overall morphologic details, compared with those provided by the other fixatives. mDF also provided excellent staining of the acrosome with periodic PAS staining and superior detect