Frederick P M, van der Molen H J, Klepper D, Galjaard H
J Cell Sci. 1977 Aug;26:339-46. doi: 10.1242/jcs.26.1.339.
The feasibility of a technique for autoradiography of diffusible substances has been further tested by analysing the localization of steroids in rat testes with the light-and electron-microscope. Testes of rats were perfused with tritiated testosterone (3 min) followed by 15-min perfusion with buffer containing a 100-fold of unlabelled testosterone. Tissue samples were frozen, freeze dried, fixed in osmium vapour and embedded in Epon. To exclude extraction of steroids, contact with water and other solvents was prevented during cutting of thin sections on an ultracryotome and further treatments for autoradiography. Light- and electron-microscopic observations indicate that the highest concentration of labelled testosterone was present within the basal parts of the Sertoli cell cytoplasm and in lipid inclusions of Sertoli cells within the seminiferous tubules. This is the first account of autoradiography of steroids at the electron-microscope level.
通过用光镜和电镜分析类固醇在大鼠睾丸中的定位，对一种用于可扩散物质放射自显影技术的可行性进行了进一步测试。给大鼠睾丸灌注氚标记的睾酮（3分钟），随后用含有100倍未标记睾酮的缓冲液灌注15分钟。将组织样本冷冻、冻干、用锇蒸汽固定并包埋在环氧树脂中。为了排除类固醇的提取，在使用超薄切片机切割薄片以及进行放射自显影的进一步处理过程中，防止与水和其他溶剂接触。光镜和电镜观察表明，标记睾酮的最高浓度存在于支持细胞胞质的基部以及生精小管内支持细胞的脂质包涵体中。这是首次在电镜水平对类固醇进行放射自显影的报道。
