Click-ExM enables expansion microscopy for all biomolecules

Nat Methods. 2021 Jan;18(1):107-113. doi: 10.1038/s41592-020-01005-2. Epub 2020 Dec 7.

Abstract

Expansion microscopy (ExM) allows super-resolution imaging on conventional fluorescence microscopes, but has been limited to proteins and nucleic acids. Here we develop click-ExM, which integrates click labeling into ExM to enable a 'one-stop-shop' method for nanoscale imaging of various types of biomolecule. By click labeling with biotin and staining with fluorescently labeled streptavidin, a large range of biomolecules can be imaged by the standard ExM procedure normally used for proteins. Using 18 clickable labels, we demonstrate click-ExM on lipids, glycans, proteins, DNA, RNA and small molecules. We demonstrate that click-ExM is applicable in cell culture systems and for tissue imaging. We further show that click-ExM is compatible with signal-amplification techniques and two-color imaging. Click-ExM thus provides a convenient and versatile method for super-resolution imaging, which may be routinely used for cell and tissue samples.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism*
  • Click Chemistry*
  • DNA / analysis
  • HeLa Cells
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Lipids / analysis
  • Macromolecular Substances / analysis*
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence / methods*
  • Myocytes, Cardiac / metabolism*
  • Organ Specificity
  • Polysaccharides / analysis
  • Proteins / analysis
  • RNA / analysis
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Lipids
  • Macromolecular Substances
  • Polysaccharides
  • Proteins
  • RNA
  • DNA