Light microscopy of proteins in their ultrastructural context

Nat Commun. 2020 Jul 31;11(1):3850. doi: 10.1038/s41467-020-17523-8.

Abstract

Resolving the distribution of specific proteins at the nanoscale in the ultrastructural context of the cell is a major challenge in fluorescence microscopy. We report the discovery of a new principle for an optical contrast equivalent to electron microscopy (EM) which reveals the ultrastructural context of the cells with a conventional confocal microscope. By decrowding the intracellular space through 13 to 21-fold physical expansion while simultaneously retaining the proteins, bulk (pan) labeling of the proteome resolves local protein densities and reveals the cellular nanoarchitecture by standard light microscopy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamides / chemistry
  • Cross-Linking Reagents / chemistry
  • Fluorescent Dyes / chemistry
  • HeLa Cells
  • Humans
  • Hydrogels / chemistry
  • Imaging, Three-Dimensional / methods*
  • Intracellular Space / chemistry
  • Microscopy, Fluorescence / methods*
  • Proteome / analysis*
  • Staining and Labeling / methods*
  • Succinimides / chemistry
  • Tissue Embedding / methods

Substances

  • Acrylamides
  • Cross-Linking Reagents
  • Fluorescent Dyes
  • Hydrogels
  • Proteome
  • Succinimides
  • N,N'-(1,2-dihydroxyethylene)bisacrylamide
  • N-hydroxysuccinimide