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

TL;DR

This paper demonstrates that infrared fluorescent proteins can serve as genetically encoded probes for stimulated Raman scattering microscopy, enabling super-multiplex cell imaging with potential for advanced biological studies.

Contribution

It introduces the use of infrared fluorescent proteins as genetically encoded SRS probes, leveraging electronic pre-resonance enhancement for improved imaging.

Findings

IRFPs enable SRS imaging of cell nuclei.

SRS imaging benefits from electronic pre-resonance with IRFPs.

This approach paves the way for Raman-based genetically encoded cell imaging.

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.