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冷冻超薄切片术及免疫细胞化学在肌肉精细结构分析中的应用

Cryoultramicrotomy and immunocytochemistry in the analysis of muscle fine structure.

作者信息

Thornell L E, Butler-Browne G S, Carlsson E, Eppenberger H M, Fürst D O, Grove B K, Holmbom B, Small J V

出版信息

Scan Electron Microsc. 1986(Pt 4):1407-18.

PMID:3544193
Abstract

Cryoultramicrotomy, which avoids the use of harsh fixation procedures, deleterious dehydration and plastic embedding can be combined with immunocytochemistry to determine the ultrastructural localization of cellular proteins. Our attempts to use the cryosectioning technique in combination with immunolabelling to bridge the gap between light and electron microscopic analysis of muscle morphology have enabled us to obtain new information on fibre typing at the ultrastructural level. Furthermore, we have obtained a marked improvement in the resolution of myofibrillar structures by using semithin cryosections for fluorescence microscopy. Data are also presented on correlated light and electron microscope immunocytochemistry of myocardial intermediate filaments confirming the presence of longitudinally oriented intermediate filaments of desmin in the region of the intercalated discs of mammalian cardiac myocytes, whereas elsewhere in the myocyte the bulk of intermediate filaments of desmin is concentrated in the intermyofibrillar space at the level of the Z disc.

摘要

冷冻超薄切片术避免了使用苛刻的固定程序、有害的脱水和塑料包埋,可与免疫细胞化学相结合来确定细胞蛋白质的超微结构定位。我们尝试将冷冻切片技术与免疫标记相结合,以弥合肌肉形态学光镜和电镜分析之间的差距,这使我们能够在超微结构水平上获得有关纤维类型的新信息。此外,通过使用半薄冷冻切片进行荧光显微镜检查,我们在肌原纤维结构的分辨率上有了显著提高。还展示了关于心肌中间丝的相关光镜和电镜免疫细胞化学数据,证实了在哺乳动物心肌细胞闰盘区域存在纵向排列的结蛋白中间丝,而在心肌细胞的其他部位,结蛋白中间丝主要集中在Z盘水平的肌原纤维间空间。

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