- Shi, Xiaoyu;
- Li, Qi;
- Dai, Zhipeng;
- Tran, Arthur A;
- Feng, Siyu;
- Ramirez, Alejandro D;
- Lin, Zixi;
- Wang, Xiaomeng;
- Chow, Tracy T;
- Chen, Jiapei;
- Kumar, Dhivya;
- McColloch, Andrew R;
- Reiter, Jeremy F;
- Huang, Eric J;
- Seiple, Ian B;
- Huang, Bo
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 superresolution stochastic optical reconstruction microscopy (STORM), we have achieved molecular resolution in the visualization of polyhedral lattice of clathrin-coated pits in situ.