Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

Stimulated Raman scattering (SRS) microscopy is a nonlinear optical technique for label-free chemical imaging. This analytical tool delivers maps at high speed, and spatial resolution of thin samples by directly interrogating their molecular vibrations. In its standard implementation, SRS narrowband forms images with only single vibrational frequency time. However, this approach not hinders the specificity but also neglects wealth information encoded within spectra. These limitations can be overcome broadband SRS, an implementation capable extracting spectrum per pixel image in parallel. hyperspectral data that, when coupled chemometric analysis, maximizes amount retrieved from specimen. Thus, improves system, allowing quantitative determination concentration different constituents sample. Here, we report protocol imaging microscopy, based on home-built microscope operating custom differential multichannel-lock-in amplifier detection. It discusses sample preparation, alignment apparatus, analysis. By acquiring spectra, illustrates how to identify species mixture, determining relative concentrations.