大鼠睾丸组织固定法的优选研究

Selection of optimal fixative for preservation of rat testis

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-Bouin's、Bouin's和改良Davidson's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Davidson's固定液较NBF-Bouin's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Davidson's固定液在需要对精子发生进行分期时,其PAS染色的效果与Bouin's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。

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 Bouin＇s fluid, Bouin＇s fluid and modified Davidson＇s fluid （mDF）, and the morphology of the testis was compared using hematoxylin and eosin （HE）-staining, periodic acidSehiff （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 seminfferous tubules, and superior overall morphologic details, compared with those provided by the other fixatives, mDF also provided excellent staining of the acrusome with periodic PAS staining and superior detection of androgen receptor with immunohistochemistry. Conclusions The results of our study demonstrate that the modified Davidson＇s fluid provides better preservation of the testis tissue and moruholo^ic details in comvarison with the rest of the fixatives.