Click-ExM enables expansion microscopy for all biomolecules
Click-ExM enables expansion microscopy for all biomolecules
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Click-ExM 可对所有生物分子进行扩展显微镜检查
DOI：
10.1038/s41592-020-01005-2
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发表时间：
2020-12-07
期刊：
影响因子：
48
通讯作者：
Chen, Xing
中科院分区：
文献类型：
--
作者：
Sun, De-en;Fan, Xinqi;Chen, Xing
扩展显微镜（ExM）允许在常规荧光显微镜上进行超分辨率成像，但仅限于蛋白质和核酸。在这里，我们开发了click-ExM，它将点击标记集成到ExM中，以实现各种类型生物分子的纳米级成像的“一站式”方法。通过用生物素点击标记和用荧光标记的链霉亲和素染色，大范围的生物分子可以通过通常用于蛋白质的标准ExM程序成像。使用18个可点击的标签，我们证明了点击ExM的脂质，聚糖，蛋白质，DNA，RNA和小分子。我们证明，点击ExM是适用于细胞培养系统和组织成像。我们进一步表明，点击ExM是兼容的信号放大技术和双色成像。因此，Click-ExM为超分辨率成像提供了一种方便和通用的方法，可常规用于细胞和组织样品。Click-ExM使用基于点击化学的标记来增加扩增显微镜的多功能性。Click-ExM可对多种生物分子进行成像，包括脂质、聚糖、蛋白质、DNA、RNA和小分子。
Expansion microscopy (ExM) allows super-resolution imaging on conventional fluorescence microscopes, but has been limited to proteins and nucleic acids. Here we develop click-ExM, which integrates click labeling into ExM to enable a 'one-stop-shop' method for nanoscale imaging of various types of biomolecule. By click labeling with biotin and staining with fluorescently labeled streptavidin, a large range of biomolecules can be imaged by the standard ExM procedure normally used for proteins. Using 18 clickable labels, we demonstrate click-ExM on lipids, glycans, proteins, DNA, RNA and small molecules. We demonstrate that click-ExM is applicable in cell culture systems and for tissue imaging. We further show that click-ExM is compatible with signal-amplification techniques and two-color imaging. Click-ExM thus provides a convenient and versatile method for super-resolution imaging, which may be routinely used for cell and tissue samples. Click-ExM uses click-chemistry-based labeling to increase the versatility of expansion microscopy. Click-ExM enables imaging of numerous classes of biomolecules including lipids, glycans, proteins, DNA, RNA and small molecules.