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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