Anatomical and chemical information can be linked by Raman imaging. Behind every pixel of the image is a Raman spectrum, which contains all the information as a molecular fingerprint. Yet to understand the spectra, the bands have to be assigned to components and their molecular structures. Although the lignin distribution is easily tracked in plant tissues, the assignment of the spectra is not good enough to allow in-depth analysis of the composition. Assignments of three lignin model compounds were derived from polarization measurements and quantum-chemical computations. Raman spectra of coniferyl alcohol crystals showed orientation dependence, which helped in band assignment. Abietin showed a Raman spectrum that was very similar to the spectrum of coniferyl alcohol, whereas its IR spectrum was very different due to bands of the sugar moiety. The Raman spectrum of coniferyl aldehyde is affected by the crystal order of molecules. All three compounds show much stronger band intensities than unconjugated single aromatic rings, indicating that the bulk of the lignin structure has significantly reduced contribution to Raman band intensities. Therefore, it is possible to highlight certain structures of lignin with Raman spectroscopy, because low amounts of a compound do not necessarily mean weak features in the spectrum.
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