Advanced ratiometric image analysis for super-resolution hyperspectral SRS microscopy

Abstract

We have developed a novel stimulated Raman scattering (SRS) imaging platform to visualize various biomolecules at a resolution beyond the traditional diffraction limit of optical microscopy. Applying a super-resolution deconvolution algorithm, Adam based Pointillism Deconvolution (A-PoD), to hyperspectral SRS images, we can measure nanoscopic distributions of these molecules in cells. In this study, we showcase the application of A-PoD for SRS images of lipids, protein, unsaturated lipid, and saturated lipids in rat olfactory bulbs under hypoxia. The analysis result shows distinct distributions of unsaturated lipid in glomeruli of olfactory bulbs in hypoxia and control rats. This approach shows the capability of revealing nanoscopic molecular composition and metabolic activities. When applied in a super-resolution context with new workflow, this approach holds significant promise for advancing early detection and understanding of diseases.