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通过超薄切片术对可扩散化合物进行电子显微镜干装放射自显影术。

Electron microscopic dry-mounting radioautography for diffusible compounds by means of ultracryotomy.

作者信息

Nagata T, Murata F

出版信息

Histochemistry. 1977 Oct 3;54(1):75-82. doi: 10.1007/BF00493331.

Abstract

Cultured HeLa cells or mouse liver and pancreas tissues were labeled with 3H-thymidine, -uridine or -glycine for varying periods in vitro, frozen in liquid nitrogen and cut on an LKB ultrotome equipped with LKB Cryokit. Dry ultrathin sections were mounted on grid meshes and were either air-dried, freeze-substituted or freeze-dried, and were covered with dry films of radioautographic emulsions, exposed, developed, stained and were observed in electron microscopes. After a number of trials, it was possible to obtain fairly good preservation of both cell structure and radioisotopes by means of freeze-dried and dry-mounted ultrathin frozen sections. However, the results are not completely satisfactory at the present time.

摘要

将培养的海拉细胞或小鼠肝脏及胰腺组织在体外以不同时间用³H-胸腺嘧啶核苷、³H-尿嘧啶核苷或³H-甘氨酸进行标记,在液氮中冷冻,并在配备LKB冷冻切片机的LKB超薄切片机上切片。干燥的超薄切片被放置在网格上,要么自然风干、冷冻置换要么冷冻干燥,然后覆盖上放射性自显影片的干膜,进行曝光、显影、染色,并在电子显微镜下观察。经过多次试验,通过冷冻干燥和干式固定的超薄冷冻切片能够较好地保存细胞结构和放射性同位素。然而,目前结果仍不完全令人满意。

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