Multicolour chemical imaging of plant tissues with hyperspectral stimulated Raman scattering microscopy

Recent development of stimulated Raman scattering (SRS) microscopy allows for label-free biological imaging with chemical specificity based on molecular-vibrational signatures. In particular, hyperspectral SRS imaging can acquire a molecular-vibrational spectrum at each pixel, allowing us not only to investigate the spectral difference of various biological molecules but also to discriminate different constituents based on their spectral difference. However, the number of constituents discriminated in previous label-free SRS imaging was limited to four because of the subtleness of spectral difference. Here, we report hyperspectral SRS imaging of plant tissues including leaves of Camellia japonica, roots of Arabidopsis thaliana, and thalli of a liverwort Marchantia polymorpha L. We show that SRS can discriminate as many as six components in Marchantia polymorpha L. without labeling. Our results demonstrate the effectiveness of hyperspectral SRS imaging as a tool for label-free multicolour imaging analysis of various biomolecules in plant tissues.

Automated summary

Stimulated Raman scattering (SRS) microscopy enables label-free biological imaging with chemical specificity and discrimination of multiple components. Hyperspectral SRS imaging can identify up to six components in plant tissues without labeling, showcasing its efficacy as a tool for label-free multicolour imaging analysis of various biomolecules.