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. 2019 Nov 4;9:15904. doi: 10.1038/s41598-019-52347-7

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© The Author(s) 2019

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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Brightfield and fluorescence microscopy analysis of safranin stained pretreated Sugarcane (SC) biomass. (a) Panel-I-III: Brightfield and fluorescence microscopy of safranin stained raw and NH₄OH pretreated SC transverse sections before and after incubation with cellulase: Panel I- Raw SC before enzyme treatment, Panel II- Raw SC and Panel III- NH₄OH pretreated SC after 120 min of cellulase treatment; Images captured for Brightfield micrograph (Column A), Green fluorescence signal (Column B) and Red fluorescence signal (Column C); Images are taken at 10X magnification; (b) Brightfield microscopy of safranin stained NaOH treated (Panel IV) and H₂SO₄ pretreated (Panel V) sugarcane biomass fraction: (i) Fibrous vascular bundle (labelled as FVB) and the fragments of surrounding parenchymatous pith cells; (ii) Individual cells of pith parenchyma separated after NaOH and H₂SO₄ pretreatments (All images at 10X magnification).