Click-ExM enables expansion microscopy for all biomolecules

Abstract We describe the development of click-expansion microscopy (Click-ExM), which integrates click-labeling into ExM to enable a“one-stop-shop” method for nanoscale imaging of various types of biomolecules. Using 18 clickable labels for click-ExM imaging of DNA, RNA, proteins, lipids, glycans and small molecules, we demonstrate its universality, compatibility with signal-amplification techniques, and broad applications in cellular and tissue imaging. The click-labeling and ExM steps could be finished within two days. This step-by-step protocol is related to the publication “Click-ExM enables expansion microscopy for all biomolecules” in Nature Methods.