切片位置对兔睾丸组织体视学研究的影响  

Comparison of the effects of embedding media and sectional locations on stereological measurement of rabbit testes

作　　者：赵圆宇[1] 王亚平[2] 郭洋[1] 张仁东[1] 杨正伟[1] 

机构地区：[1]川北医学院形态定量研究室,南充637007 [2]重庆医科大学干细胞与组织工程研究室,重庆400016

出　　处：《中国体视学与图像分析》2010年第4期428-431,共4页Chinese Journal of Stereology and Image Analysis

基　　金：四川省青年科技基金(04ZQ026-025)

摘　　要：目的比较不同包埋介质内切片位置对成年新西兰大耳白兔睾丸组织体视学研究的影响。方法将成年雄性新西兰大耳白兔麻醉后,随机抽选一侧睾丸经Bouin液固定后从中央及两端(垂直于睾丸中心长轴)切取共三个平行组织薄片。将每个组织薄片平分为二,再随机将其中一块平分得到2个(1/4圆)组织块,分别随机采用羟乙基甲基丙烯酸树脂或石蜡包埋、切片和染色后,在低倍镜(10×)下分别对睾丸组织树脂切片(25μm)和石蜡切片(7μm)不同切片位置进行观察:紧贴白膜选择若干视野(所有视野间无任何重叠),再向扇形切片中心连续选择3个视野,利用体视学方法估计各视野睾丸组织内生精小管的体积分数。结果石蜡切片较树脂切片估计的生精小管体积分数减少15.9%;树脂切片不同部位估计的生精小管体积分数无差异,而石蜡切片不同部位估计的结果存在差异:距离白膜愈远,体积分数愈小,近切片中心与近白膜相比显著减少了30.1%。结论石蜡包埋能引起睾丸组织不均匀皱缩,因此,当应用石蜡切片进行睾丸体积分数研究时,应考虑不均匀组织皱缩因素。Objective To compare the influence of two embedding media and sectional sites on stereological measurement of rabbit testes.Methods Each testis was taken and fixed in Bouin＇s fixative from each anaesthetized adult male rabbit.The left or right testis was selected randomly.Then,three parallel tissue slices from the middle and the two poles of the testis were obtained（perpendicular to the longitudinal axis of the testis）.Each slice was divided equally into two pieces.One piece was selected randomly and divided equally again.The two blocks were embedded in paraffin or methacrylate resin,respectively.One section from each block was cut and stained.Paraffin sections of 7 mm thickness and methacrylate sections of 25 mm thickness were observed and measured on a computer screen using a 10 lens.Several counting areas were selected from sub-albuginea and there was not any overlap between these areas.Then three fields towards the center of the fan-shaped section was sampled continuously.The volume fraction of the seminiferous tubules was estimated with the stereological method.Results The volume fraction of the seminiferous tubules in paraffin sections was decreased by 15.9% in comparison with that in methacrylate sections.There was no significant difference of the volume fraction of the seminiferous tubules in different sampled sites of methacrylate sections.On contrast,there was a significant difference of the volume fraction of the seminiferous tubules in different sampled sites of paraffin sections.Compared with the mean volume fraction of seminiferous tubules in sub-albuginea,the volume fraction of seminiferous tubules near the section center was decreased by 30.1%.Conclusions The non-uniform shrinkage caused by paraffin embedding should be taken into account when the volume fraction of seminiferous tubules in testis tissue is estimated from paraffin sections.

关 键 词：生精小管 切片位置 甲基丙烯酸树脂 石蜡 体视学 

分 类 号：Q954.6[生物学—动物学]