Fixation-resistant photoactivatable fluorescent proteins for CLEM

Nat Methods. 2015 Mar;12(3):215-8, 4 p following 218. doi: 10.1038/nmeth.3225. Epub 2015 Jan 12.

Abstract

Fluorescent proteins facilitate a variety of imaging paradigms in live and fixed samples. However, they lose their fluorescence after heavy fixation, hindering applications such as correlative light and electron microscopy (CLEM). Here we report engineered variants of the photoconvertible Eos fluorescent protein that fluoresce and photoconvert normally in heavily fixed (0.5-1% OsO4), plastic resin-embedded samples, enabling correlative super-resolution fluorescence imaging and high-quality electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • CHO Cells
  • Cricetulus
  • Fluorescence
  • HeLa Cells
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Microscopy, Electron, Scanning / methods*
  • Microscopy, Electron, Transmission / methods*
  • Molecular Imaging / methods
  • Molecular Sequence Data
  • Osmium Tetroxide / chemistry
  • Photochemistry / methods
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Osmium Tetroxide