College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
Beijing National Laboratory for Molecular Sciences, Peking University, Beijing, China.
Nat Methods. 2021 Jan;18(1):107-113. doi: 10.1038/s41592-020-01005-2. Epub 2020 Dec 7.
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.
扩展显微镜（ExM）允许在传统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，我们开发了点击扩展显微镜（click-ExM），它将点击标记整合到 ExM 中，为各种类型的生物分子的纳米级成像提供了一种“一站式”方法。通过使用生物素进行点击标记，并使用荧光标记的链霉亲和素进行染色，可以通过通常用于蛋白质的标准 ExM 程序对大量生物分子进行成像。使用 18 种可点击的标签，我们在脂质、聚糖、蛋白质、DNA、RNA 和小分子上展示了 click-ExM。我们证明了 click-ExM 适用于细胞培养系统和组织成像。我们进一步表明，click-ExM 与信号放大技术和双色成像兼容。因此，click-ExM 为超分辨率成像提供了一种方便、通用的方法，可常规用于细胞和组织样本。
