用于蛋白质原位定位的相关显微镜技术：使用氟纳米金、金增强和低温树脂的包埋前免疫电子显微镜技术
Correlative Microscopy for Localization of Proteins In Situ: Pre-embedding Immuno-Electron Microscopy Using FluoroNanogold, Gold Enhancement, and Low-Temperature Resin.
作者信息
Boassa Daniela
机构信息
National Center for Microscopy and Imaging Research, University of California San Diego, La Jolla, CA, 92093, USA,
出版信息
Methods Mol Biol. 2015;1318:173-80. doi: 10.1007/978-1-4939-2742-5_17.
Immuno-electron microscopy (immuno-EM) is a technique that has been used widely to determine subcellular localization of proteins. Different approaches are available for immuno-EM: pre-embedding method, post-embedding, and cryosectioning (Tokuyasu "style"). Here we describe a pre-embedding technique that allows the labeling of a target protein in situ, retention of fluorescence signal in plastic, and its localization at the EM level in a given cellular context. The procedure can be technically challenging and labor intensive: it requires optimization of fixation protocols to better preserve the cellular morphology and screening of compatible antibodies. Nevertheless, immuno-EM can be a powerful localization tool.
免疫电子显微镜技术（免疫电镜）是一种广泛用于确定蛋白质亚细胞定位的技术。免疫电镜有多种方法：包埋前法、包埋后法和冷冻切片法（德康安 “式”）。在此，我们描述一种包埋前技术，该技术可对原位目标蛋白进行标记，在塑料中保留荧光信号，并在给定的细胞环境中在电子显微镜水平上确定其定位。该过程在技术上具有挑战性且劳动强度大：它需要优化固定方案以更好地保存细胞形态，并筛选兼容的抗体。尽管如此，免疫电镜仍是一种强大的定位工具。
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