PT - JOURNAL ARTICLE AU - Xiaoyu Shi AU - Qi Li AU - Zhipeng Dai AU - Arthur A. Tran AU - Siyu Feng AU - Alejandro D. Ramirez AU - Zixi Lin AU - Xiaomeng Wang AU - Tracy T. Chow AU - Ian B. Seiple AU - Bo Huang TI - Label-retention expansion microscopy AID - 10.1101/687954 DP - 2019 Jan 01 TA - bioRxiv PG - 687954 4099 - http://biorxiv.org/content/early/2019/07/02/687954.short 4100 - http://biorxiv.org/content/early/2019/07/02/687954.full AB - Expansion microscopy (ExM) improves the resolution of fluorescence microscopy by physically expanding the sample embedded in a hydrogel1–4. Since its invention, ExM has been successfully applied to a wide range of cell, tissue and animal samples 2–9. 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 protein labeling using enzymatic 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 5 nm resolution in the visualization of polyhedral lattice of clathrin-coated pits in situ.