An experimental evaluation of the information content of two complimentary techniques, linear Raman and coherent anti-Stokes Raman scattering (CARS) microscopy, is presented. CARS is a nonlinear variant of Raman spectroscopy that enables rapid acquisition of images within seconds in combination with laser scanning microscopes. CARS images were recorded from thin colon tissue sections at 2850, 1660, 1450 and 1000 cm(-1) and compared with Raman images. Raman images were obtained from univariate and multivariate (k-means clustering) methods, whereas all CARS images represent univariate results. Variances within tissue sections could be visualized in chemical maps of CARS and Raman images. However, identification of tissue types and characterization of variances between different tissue sections were only possible by analysis of cluster mean spectra, obtained from k-means cluster analysis. This first comparison establishes the foundation for further development of the CARS technology to assess tissue.
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