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

Lucie Ludvikova; Emma Simon; Mathieu Deygas; Thomas Panier; Marie-Aude Plamont; Jean Ollion; Alison Tebo; Matthieu Piel; Ludovic Jullien; Lydia Robert; Thomas Le Saux; Agathe Espagne

doi:10.1117/12.3008062

13 March 2024 Near-infrared co-illumination of fluorescent proteins reduces photobleaching and phototoxicity

Lucie Ludvikova, Emma Simon, Mathieu Deygas, Thomas Panier, Marie-Aude Plamont, Jean Ollion, Alison Tebo, Matthieu Piel, Ludovic Jullien, Lydia Robert, Thomas Le Saux, Agathe Espagne

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Proceedings Volume PC12862, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications XV; PC128620K (2024) https://doi.org/10.1117/12.3008062
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

We will present a new method to reduce the photobleaching of fluorescent proteins and the associated phototoxicity. Our method exploits a photophysical process known as reverse intersystem crossing, which we induce by near-infrared co-illumination during fluorophore excitation. This dual illumination method typically reduces photobleaching effects 4-fold, can be easily implemented on commercial microscopes and is effective in eukaryotic and prokaryotic cells with a wide range of fluorescent proteins.

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Lucie Ludvikova, Emma Simon, Mathieu Deygas, Thomas Panier, Marie-Aude Plamont, Jean Ollion, Alison Tebo, Matthieu Piel, Ludovic Jullien, Lydia Robert, Thomas Le Saux, and Agathe Espagne "Near-infrared co-illumination of fluorescent proteins reduces photobleaching and phototoxicity", Proc. SPIE PC12862, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications XV, PC128620K (13 March 2024); https://doi.org/10.1117/12.3008062

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KEYWORDS

Fluorescent proteins

Photobleaching

Light sources and illumination

Fluorophores

Bacteria

Fluorescence imaging

In vivo imaging

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