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

Regan, David ORCID: https://orcid.org/0000-0003-0420-3481, Aksakal, Ozan, Zitti, Athena, McLarnon, John, Lipka-Lloyd, Magdalena, Rizkallah, Pierre J. ORCID: https://orcid.org/0000-0002-9290-0369, Warren, Anna J., Watson, Peter D. ORCID: https://orcid.org/0000-0003-0250-7852, Langbein, Wolfgang ORCID: https://orcid.org/0000-0001-9786-1023, Jones, D. Dafydd ORCID: https://orcid.org/0000-0001-7709-3995 and Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 2025. Genetically encoding stimulated Raman-scattering probes for cell imaging using infrared fluorescent proteins. arXiv. 10.48550/arXiv.2511.08564

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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.

Item Type:	Working paper
Date Type:	Published Online
Status:	Submitted
Schools:	Schools > Biosciences Schools > Physics and Astronomy
Publisher:	arXiv
Last Modified:	03 Feb 2026 09:58
URI:	https://orca.cardiff.ac.uk/id/eprint/184364

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