A novel optimized pre-embedding antibody-labelling correlative light electron microscopy technique.

Access microbiology Pub Date : 2024-02-20 eCollection Date: 2024-01-01 DOI:10.1099/acmi.0.000750.v3
Nicole Doyle, Jennifer Simpson, Philippa C Hawes, Helena J Maier
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Abstract

In the intricate environment of a cell, many studies seek to discover the location of specific events or objects of interest. Advances in microscopy in recent years have allowed for high detail views of specific areas of cells of interest using correlative light electron microscopy (CLEM). While this powerful technique allows for the correlation of a specific area of fluorescence on a confocal microscope with that same area in an electron microscope, it is most often used to study tagged proteins of interest. This method adapts the correlative method for use with antibody labelling. We have shown that some cellular structures are more sensitive than others to this process and that this can be a useful technique for laboratories where tagged proteins or viruses, or dedicated CLEM instruments are not available.

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新型优化预包埋抗体标记相关光电子显微镜技术。
在错综复杂的细胞环境中,许多研究都试图发现特定事件或感兴趣对象的位置。近年来,显微镜技术的进步使得人们可以利用相关光电子显微镜(CLEM)对感兴趣的细胞特定区域进行高细节观察。虽然这种强大的技术可以将共聚焦显微镜上特定区域的荧光与电子显微镜中相同区域的荧光相关联,但它最常用于研究感兴趣的标记蛋白质。这种方法将相关法与抗体标记法相结合。我们已经证明,某些细胞结构对这一过程比其他结构更敏感,对于没有标记蛋白质或病毒或专用 CLEM 仪器的实验室来说,这可能是一项有用的技术。
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