Label-retention expansion microscopy

Abstract 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 polymerizat...

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Bibliographic Details
Published inbioRxiv
Main Authors 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
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 04.03.2021
Subjects
Clathrin
Coated pits
Hydrogels
Microscopy
Polymerization
Stochasticity
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Summary:Abstract 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 Statement The authors have declared no competing interest. Footnotes * New data on brain slice and basal body imaging
DOI:10.1101/687954