Immunogold labelling for scanning electron microscopy.
Immunogold labelling for scanning electron microscopy.
复制标题
用于扫描电子显微镜的免疫金标记。
DOI：
10.1007/978-1-60761-783-9_24
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发表时间：
2010
影响因子：
--
通讯作者：
J. Fišerová
中科院分区：
文献类型：
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作者：
M. Goldberg;J. Fišerová
扫描电子显微镜是有用的生物工具，可用于对整个生物体、组织、细胞、细胞成分和大分子的表面进行成像。表面上存在的过程和结构可以以大约 x10 到 x1,000,000 的放大倍数以伪或真实 3D 方式成像。因此，可以看到整个多细胞生物体，例如苍蝇，或嵌入其细胞膜之一的单个蛋白质。为了以高分辨率识别该蛋白质，或者以较低的放大倍数观察和量化其分布，可以用抗体标记样品。任何可能暴露的表面都可以通过这种方式进行研究。这里介绍的是扫描电子显微镜免疫金标记的通用方法，使用两个样本示例：分离的核膜和哺乳动物培养细胞的细胞骨架。讨论了样品制备、固定、免疫金标记、干燥、金属涂层和成像的各种参数，以便从不同类型的标本中获得最佳的免疫金扫描电子显微镜结果。
Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo or real 3D at magnifications of anything from about x10 to x1,000,000. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualised. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labelled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labelling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labelling, drying, metal coating and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.