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arXiv · 2511.08564

Genetically encoding stimulated Raman-scattering probes for cell imaging using infrared fluorescent proteins

David Regan·Ozan Aksakal·Athena Zitti·John McLarnon·Magdalena Lipka-Lloyd·Pierre J. Rizkallah·Anna J. Warren·Peter D. Watson·Wolfgang Langbein·D. Dafydd Jones·Paola Borri
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Abstract

Stimulated Raman scattering (SRS) microscopy offers great potential to surpass fluorescent-based approaches, owing to the sharp linewidth of Raman vibrations amenable to super-multiplex cell imaging, but currently lacks one crucial component: genetically encodable tags equivalent to fluorescent proteins. Here, we show that infrared fluorescent proteins (IRFPs) can be used as genetically encoded SRS probes and benefit from the electronic pre-resonant SRS enhancement effect with near-infrared exciting pulses, comparable to synthetic dyes reported in the literature. SRS imaging of the nucleus in mammalian cells is demonstrated where a histone protein is fused to an IRFP. This work opens the route towards Raman-based cell imaging using genetically encoded probes, motivating efforts in solving the challenges of photostability and creating a vibrational palette.

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David Regan, Ozan Aksakal, Athena Zitti, John McLarnon, Magdalena Lipka-Lloyd, Pierre J. Rizkallah, Anna J. Warren, Peter D. Watson, Wolfgang Langbein, D. Dafydd Jones, Paola Borri. 2025-11-11. Genetically encoding stimulated Raman-scattering probes for cell imaging using infrared fluorescent proteins. https://doi.org/10.1021/acs.jpclett.6c02085

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