Click-ExM 使所有生物分子的扩展显微镜成为可能。

Click-ExM enables expansion microscopy for all biomolecules.

机构信息

College of Chemistry and Molecular Engineering, Peking University, Beijing, China.

Beijing National Laboratory for Molecular Sciences, Peking University, Beijing, China.

出版信息

Nat Methods. 2021 Jan;18(1):107-113. doi: 10.1038/s41592-020-01005-2. Epub 2020 Dec 7.

DOI:10.1038/s41592-020-01005-2

PMID:33288959

Abstract

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.

摘要

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。