Stimulated Raman Scattering Imaging Techniques in Biological Systems

Stimulated Raman scattering (SRS) microscopy has emerged as a powerful label-free imaging modality that exploits intrinsic molecular vibrations to generate high-contrast chemical maps of biological specimens. By synchronously driving vibrational modes with two laser beams—the pump and the Stokes—SRS achieves orders of magnitude greater sensitivity and speed than spontaneous Raman methods, enabling real-time visualisation of cellular metabolites, lipids, proteins and nucleic acids. Recent technical innovations span both hardware and computational domains: time-domain strategies circumvent the time-energy uncertainty limit to deliver natural-linewidth spectral resolution; hyperspectral acquisition modes coupled with advanced unmixing algorithms facilitate multiplexed mapping of biomolecular subtypes; and machine-learning-driven volumetric reconstructions push the boundaries of imaging depth, speed and fidelity. These advances have been applied to a wide range of biological systems, from tracking de novo lipogenesis in single cancer cells to rapid intraoperative histology and in vivo metabolic studies. Together, they underline the global significance of SRS as a non-invasive, quantitative tool for probing physiological and pathological processes at subcellular resolution.

Research from Nature Portfolio

A hyperspectral SRS platform combined with a penalised reference matching algorithm has been used to distinguish multiple lipid subtypes in diverse biological specimens. This approach achieves subcellular resolution and rapid data processing, revealing distinct spatial distributions of high-density lipoprotein in human kidney, lipid ratio variations in mouse hippocampus, and sphingosine and cardiolipin localisation in human brain tissue. The integration of computational spectral unmixing with broad-band SRS imaging enhances chemical specificity and throughput, enabling detailed studies of lipid metabolism across organs and species.

Building on vibrational contrast and stable isotope labelling, another study introduced deuterium oxide probing with in situ SRS microscopy to monitor metabolic dynamics in living animals. By mapping carbon–deuterium bonds and applying spectral unmixing, researchers could image lipid biosynthesis, protein turnover and DNA synthesis in intact tissues. This non-invasive technique provides macromolecular-selective metabolic readouts, offering a versatile platform for investigations of development, tissue homeostasis, ageing and tumour heterogeneity.

Stimulated Raman Scattering Imaging Techniques in Biological Systems publication trend

The graph below shows the total number of articles in stimulated raman scattering imaging techniques in biological systems across all publications each year (not limited to Nature Index journals).

Technical terms

Stimulated Raman Scattering (SRS): A nonlinear optical process that amplifies weak Raman signals by coherently driving vibrational modes with two synchronised lasers, yielding fast, label-free chemical imaging.

Hyperspectral imaging: Acquisition of images across a continuous spectrum of wavelengths or wavenumbers to resolve and quantify multiple chemical species within a sample simultaneously.

Penalised Reference Matching (PRM): A spectral unmixing algorithm that incorporates penalty functions to improve discrimination of overlapping Raman signatures in hyperspectral datasets.

Bioorthogonal tag: A chemically inert functional group introduced into biomolecules that produces a distinct Raman signal without perturbing native cellular processes.

Quantum coherence: The maintenance of well-defined phase relationships between quantum states, employed in time-domain SRS to manipulate and decode vibrational wave-packet dynamics.

References

1. Computational coherent Raman scattering imaging: breaking physical barriers by fusion of advanced instrumentation and data science. eLight (2023).
2. Transient stimulated Raman scattering spectroscopy and imaging. Light: Science & Applications (2024).
3. Multi-molecular hyperspectral PRM-SRS microscopy. Nature Communications (2024).
4. Optical imaging of metabolic dynamics in animals. Nature Communications (2018).
5. Direct Visualization of De novo Lipogenesis in Single Living Cells. Scientific Reports (2014).
6. Rapid histology of laryngeal squamous cell carcinoma with deep-learning based stimulated Raman scattering microscopy. Theranostics (2019).