Near-infrared co-illumination of fluorescent proteins reduces photobleaching and phototoxicity

Nat Biotechnol. 2024 Jun;42(6):872-876. doi: 10.1038/s41587-023-01893-7. Epub 2023 Aug 3.

Abstract

Here we present a method to reduce the photobleaching of fluorescent proteins and the associated phototoxicity. It exploits a photophysical process known as reverse intersystem crossing, which we induce by near-infrared co-illumination during fluorophore excitation. This dual illumination method reduces photobleaching effects 1.5-9.2-fold, can be easily implemented on commercial microscopes and is effective in eukaryotic and prokaryotic cells with a wide range of fluorescent proteins.

MeSH terms

  • Humans
  • Infrared Rays
  • Luminescent Proteins* / metabolism
  • Microscopy, Fluorescence / methods
  • Photobleaching*

Substances

  • Luminescent Proteins