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胶体金标记系统综述。

A review of the colloidal gold marker system.

作者信息

Goodman S L, Hodges G M, Livingston D C

出版信息

Scan Electron Microsc. 1980(Pt 2):133-46.

PMID:6999596
Abstract

Colloidal gold can be used as a particulate marker for the detection and localization of target molecules by various modes of microscopy (light and fluorescent microscopy, scanning and transmission electron microscopy) using both direct and indirect labeling approaches. Several techniques are available for the preparation of gold markers in a size range of 5nm to 150nm, their mean size and shape characteristics and absorption spectra varying with particle size. Under appropriate conditions, colloidal gold will bind macromolecules by non-covalent electrostatic adsorption with little change in the specific activity of the bound macromolecule. This interaction is influenced by a number of factors including ionic concentration, pH conditions (in correlation with the protein pI values) and protein stabilizing levels. Presence of reactive protein on probes can be demonstrated and quantitated by direct and indirect radioactive binding assays and agglutination assays. These assays provide convenient procedures for characterizing stability, and behaviour in storage, of gold probes. Stability of gold probes under conditons where competing proteins are present, under freeze-thaw cycles and under SEM preparation conditions have been evaluated in this paper. Some of the basic procedures in the application of gold probes to cell labeling are briefly discussed together with certain limitations of the colloidal gold marker system. A bibliography of gold probe cell labeling studies is included.

摘要

胶体金可作为一种颗粒标记物,通过各种显微镜技术(光学显微镜和荧光显微镜、扫描电子显微镜和透射电子显微镜),采用直接和间接标记方法,用于检测和定位目标分子。有几种技术可用于制备尺寸范围在5纳米至150纳米的金标记物,其平均尺寸、形状特征和吸收光谱随颗粒大小而变化。在适当条件下,胶体金将通过非共价静电吸附结合大分子,而结合的大分子的比活性变化很小。这种相互作用受多种因素影响,包括离子浓度、pH条件(与蛋白质的等电点值相关)和蛋白质稳定水平。探针上反应性蛋白质的存在可通过直接和间接放射性结合测定以及凝集测定来证明和定量。这些测定为表征金探针的稳定性以及储存行为提供了方便的程序。本文评估了在存在竞争蛋白质的条件下、冻融循环条件下以及扫描电子显微镜制备条件下金探针的稳定性。简要讨论了金探针应用于细胞标记的一些基本程序以及胶体金标记系统的某些局限性。文中还包含了金探针细胞标记研究的参考文献目录。

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