Near-infrared co-illumination of fluorescent proteins reduces photobleaching and phototoxicity
Lucie Ludvíková, Emma Simon, Mathieu Deygas, Thomas Panier, Marie‐Aude Plamont, Jean Ollion, Alison G. Tebo, Matthieu Piel, Ludovic Jullien, Lydia Robert, Thomas Le Saux, Agathe Espagne
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.
Topics & Concepts
PhotobleachingPhototoxicityFluorophoreFluorescenceIntersystem crossingChemistryBiophysicsPhotochemistryInfraredFluorescent proteinMicroscopeFluorescent labellingGreen fluorescent proteinOpticsExcited stateBiochemistryBiologyIn vitroPhysicsGeneSinglet stateNuclear physicsAdvanced Fluorescence Microscopy TechniquesCell Image Analysis Techniquesbioluminescence and chemiluminescence research