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使用免疫标记树脂切片的相关光镜和电镜检查。

Correlative light and electron microscopy using immunolabeled resin sections.

作者信息

Schwarz Heinz, Humbel Bruno M

机构信息

Max-Planck-Institut für Entwicklungsbiologie, Tuebingen, Germany.

出版信息

Methods Mol Biol. 2007;369:229-56. doi: 10.1007/978-1-59745-294-6_12.

Abstract

In correlative microscopy, light microscopy provides the overview and orientation in the complex cells and tissue, whereas electron microscopy offers the detailed localization and correlation to subcellular structures. In this chapter, we offer the detailed high-quality electron microscopical preparation methods for the optimum preservation of the cellular ultrastructure. From such preparations, serial thin sections are collected and used for comparative histochemical, immunofluorescence, and immunogold staining. In light microscopy, histological stains are used to identify the orientation of the sample, and immunofluorescence labeling is used to identify the region of interest, namely, the labeled cells expressing the macromolecule under investigation. Subsequent sections, labeled with immunogold, are analyzed by electron microscopy to identify the label within the cellular architecture at high resolution.

摘要

在相关显微镜检查中,光学显微镜提供复杂细胞和组织的整体概览与定位,而电子显微镜则能实现亚细胞结构的详细定位及关联。在本章中,我们提供详细的高质量电子显微镜制备方法,以实现细胞超微结构的最佳保存。从这些制备物中收集连续薄切片,并用于比较组织化学、免疫荧光和免疫金染色。在光学显微镜下,组织学染色用于确定样本的方向,免疫荧光标记用于识别感兴趣的区域,即表达所研究大分子的标记细胞。随后用免疫金标记的切片通过电子显微镜进行分析,以高分辨率识别细胞结构内的标记物。

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