RT Journal Article SR Electronic T1 Label-retention expansion microscopy JF bioRxiv FD Cold Spring Harbor Laboratory SP 687954 DO 10.1101/687954 A1 Xiaoyu Shi A1 Qi Li A1 Zhipeng Dai A1 Arthur A. Tran A1 Siyu Feng A1 Alejandro D. Ramirez A1 Zixi Lin A1 Xiaomeng Wang A1 Tracy T. Chow A1 Jiapei Chen A1 Dhivya Kumar A1 Andrew R. McColloch A1 Jeremy F. Reiter A1 Eric J. Huang A1 Ian B. Seiple A1 Bo Huang YR 2021 UL http://biorxiv.org/content/early/2021/03/04/687954.abstract AB Expansion microscopy (ExM) increases the effective resolving power of any microscope by expanding the sample with swellable hydrogel. Since its invention, ExM has been successfully applied to a wide range of cell, tissue and animal samples. Still, fluorescence signal loss during polymerization and digestion limits molecular-scale imaging using ExM. Here we report the development of label-retention ExM (LR-ExM) with a set of trifunctional anchors that not only prevent signal loss but also enable high-efficiency labeling using SNAP and CLIP tags. We have demonstrated multicolor LR-ExM for a variety of subcellular structures. Combining LR-ExM with super-resolution Stochastic Optical Reconstruction Microscopy (STORM), we have achieved molecular resolution in the visualization of polyhedral lattice of clathrin-coated pits in situ.Competing Interest StatementThe authors have declared no competing interest.