Deconstruction of Obscure Features in SVD-Decomposed Raman Images from P. chrysogenum Reveals Complex Mixing of Spectra from Five Cellular Constituents.

Raman imaging has transcended in recent times from being an analytical tool to a molecular profiling technique. Biomedical applications of this technique often rely on singular-value decomposition (SVD), principal component analysis (PCA), etc. for data analysis. These methods, however, obliterate the molecular information contained in the original Raman data leading to speculative interpretations based on relative intensities. In the present study, SVD analysis of the Raman images from Penicillium chrysogenum resulted in 11 spectral components and corresponding images with highly distorted spectral features and complex image contrast, respectively. To interpret the SVD results in molecular terms, we have developed a combined multivariate approach. By applying this methodology, we have successfully extracted the contribution of five biomolecular constituents of the P. chrysogenum filamentous cell to the SVD vectors. Molecular interpretability will help SVD/PCA surpass the realm of variance-based classification to a more meaningful molecular domain.