采用卵巢冰冻切片和改进H&E染色方法分析小鼠发情周期卵泡形态（英文）

Frozen section with improved H&E staining for follicular morphometric analysis of mouse ovary in oestrus cycle

通过观察卵巢组织中不同发育阶段的各级卵泡的形态和数量,并结合促性腺激素和性激素的测定能够更好地评估卵巢功能。本研究目的在于改进现有的卵巢组织切片及染色方法,建立更快观察和评价卵巢组织中各级卵泡数量和质量的方法。取动情前期、动情期、动情后期和动情间期雌性C57BL/6J小鼠卵巢,用4%多聚甲醛固定,梯度蔗糖脱水,OCT(optimal cutting temperature compound)包埋,冰冻切片(厚7μm),快速H&E染色后进行观察。结果显示,本方法能够分辨次级卵泡、窦前、窦状和排卵前卵泡,虽然不能区分和分辨始基卵泡和初级卵泡,但是其它观察结果与复杂的方法相当。以上结果提示,快速和改进的小鼠卵巢冰冻切片和H&E染色方法可以与激素分析结合,用于小鼠模型中的卵巢发育和功能研究。

Careful analysis of follicular morphology and numbers of different follicular maturation stages, in combination with the measurements of gonadotropic and sex hormone profiles, provide an accurate and rapid evaluation system of the ovarian function. The aim of this study is to improve the existing methods of ovarian tissue section preparation and staining methods, and to establish a fast and easy method to observe and evaluate follicular maturation stage and numbers using mouse ovary samples. Ovaries were collected at menstrual phases of proestrus, oestrus, metestrus and diestrus from C57BL/6J female mice. Then the ovaries were fixed in 4% paraformaldehyde, dehydrated with graded sucrose, embedded with OCT, frozen-sectioned at 7 μm thick, and subjected to quick haematoxylin ＆ eosin staining for observation. The results showed that the present method was able to distinguish secondary, preantral, antral, and preovulatory follicles. Although our method was unable to discriminate and distinguish primordial follicles and primary follicles, the results were comparable to those from more complicated techniques. We conclude that this improved and quick method can be used in combination with hormone analysis to investigate ovarian development and function in different mouse models.