Mahta Mohamadiaza, Naser Farrokhi, Pär K Ingvarsson, Asadollah Ahmadikhah
Raman spectroscopy (RS) is a rapid, label-free, and non-destructive technique for analyzing molecular and spectral characteristics of biological samples. Here, we present a reproducible protocol for glycerol-based sample preparation and RS analysis of long-term air-dried rice straw. We describe steps for preparing samples, acquiring spectral data, and preprocessing data. Our workflow enables reproducible spectral acquisition and semi-quantitative assessment of relative Raman spectral markers associated with major plant cell wall components using standard laboratory procedures. Spectral preprocessing and analysis can be performed with commercially available software, including MATLAB and OriginPro.