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

Ludvikova, Lucie; Simon, Emma; Deygas, Mathieu; Panier, Thomas; Plamont, Marie-Aude; Ollion, Jean; Tebo, Alison; Piel, Matthieu et al. · Nat Biotechnol · 2024

basic_science · Level V

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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.

Medical subject headings