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. 2018 Feb 9;8:2691. doi: 10.1038/s41598-018-20928-7

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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(a) Micrograph of toluidine blue stained cross section of the wildtype sorghum coleoptile. (b) Visible image from Raman microscopy of the central area of the section showing the vascular bundle region with the investigated area of 42 × 24 μm as marked on panel a. (c) The KMC results, with (d) the average Raman spectra for the main classes, including highest (green), medium (blue) and lowest (grey green) dhurrin content. Raman intensities were normalized for C-H stretching region (2800–3050 cm⁻¹) in order to compare the dhurrin content in different classes. (e–h) Integration Raman maps obtained with a sampling density of 1 µm were made for the following: (e) C-H stretching band in the region of approx. 2800‒3050 cm⁻¹ showing organic constituents present; (f–h) the strongest bands of the dhurrin observed at 867, 1613 and 2249 cm⁻¹, respectively. The yellow colour corresponds to the highest relative intensity of the integrated band. All material was prepared using cryosections.