Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging

This protocol compares the two most widely-used hyperspectral coherent Raman scattering techniques including the coherent anti-Stokes Raman scattering and the stimulated Raman scattering processes and this allows the biologists to choose the best imaging modality for their biological applications. Using the intrinsic vibrational signatures, coherent Raman spectroscopy is able to map out chemical information and biological samples without the need for exogenous labeling. Coherent Raman microscopy has expanded our understanding of cellular metabolism, mapping, drug distributions, disease diagnostics, and quantifying chemical changes.

This is a technique that requires sufficient training in both optics and spectroscopy. I would recommend reading a few review articles and being trained by an expert before using this technique. To begin, prepare the imaging slides by placing a piece of double-sided tape onto a cover slip and cut out a small rectangular shape of the tape from the middle of the tape to create an open area for the sample to be placed.