一种用于制备用于透射电子显微镜的分离卵巢滤泡的简单而有效的方法。
A simple and efficient method for preparation of isolated ovarian follicles for transmission electron microscopy.
机构信息
Department of OB-GYN, Women's Health Institute, Cleveland Clinic Foundation, Cleveland, OH, USA.
出版信息
J Assist Reprod Genet. 2010 Feb;27(2-3):97-101. doi: 10.1007/s10815-010-9389-4. Epub 2010 Feb 6.
OBJECTIVE
A simple method for preparation of isolated ovarian follicles for transmission electron microscopy (TEM) using transwell inserts is described.
MATERIALS AND METHODS
Pre-antral follicles were enzymatically isolated from mouse ovaries and cultured overnight on transwell insert polyester membranes. The following day, isolated ovarian follicles were processed for TEM by moving the transwell insert through successive wells containing the fixation and embedding reagents. After polymerization of the resin, the polyester membrane with the follicles embedded in the resin was disengaged from the transwell unit. The resin was sectioned. Semi-thin sections were stained with toluidine blue while ultra-thin sections were stained by uranyl acetate and examined by light microscopy and TEM, respectively.
RESULTS
Isolated ovarian follicles were easily processed in groups for TEM. Follicles were well embedded and there appeared to be no loss of tissue during processing. The ultra-structure of processed isolated ovarian follicles was well preserved with little evidence of processing artifacts.
CONCLUSIONS
In situ processing and preparation of isolated ovarian follicles by first allowing attachment on transwell insert membranes was shown to be a simple, rapid and effective method for TEM.
目的
描述一种使用 Transwell 插入物分离卵巢卵泡进行透射电镜（TEM）的简单方法。
材料与方法
从小鼠卵巢中酶解分离出原始卵泡，并在 Transwell 插入式聚酯膜上培养过夜。次日，通过将 Transwell 插入物移至含有固定和包埋试剂的连续孔中，对分离的卵巢卵泡进行 TEM 处理。树脂聚合后，将包埋在树脂中的聚酯膜与 Transwell 单元分离。然后对树脂进行切片，甲苯胺蓝染色半薄切片，醋酸铀酰染色超薄切片，分别用光镜和 TEM 观察。
结果
可轻松地将分离的卵巢卵泡分组进行 TEM 处理。卵泡包埋良好，在处理过程中似乎没有组织丢失。处理后的分离卵巢卵泡的超微结构保存良好，几乎没有处理伪影的证据。
结论
通过首先允许在 Transwell 插入式膜上附着来原位处理和制备分离的卵巢卵泡，是一种简单、快速和有效的 TEM 方法。
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