Abstract
Biomass exhibits structural and chemical complexity over multiple size scales, presenting many challenges to the effective characterization of these materials. The macroscopic nature of plants requires that some form of size reduction, such as dissection and microtomy, be performed to prepare samples and reveal features of interest for any microscopic and nanoscopic analyses. These size reduction techniques, particularly sectioning and microtomy, are complicated by the inherent porosity of plant tissue that often necessitates fixation and embedding in a supporting matrix to preserve structural integrity. The chemical structure of plant cell walls is vastly different from that of the membrane bound organelles and protein macromolecular complexes within the cytosol, which are the focus of many traditional transmission electron microscopy (TEM) investigations in structural biology; thus, staining procedures developed for the latter are not optimized for biomass. While the moisture content of biomass is dramatically reduced compared to the living plant tissue, the residual water is still problematic for microscopic techniques conducted under vacuum such as scanning electron microscopy (SEM). This requires that samples must be carefully dehydrated or that the instrument must be operated in an environmental mode to accommodate the presence of water. In this chapter we highlight tools and techniques that have been successfully used to address these challenges and present procedural details regarding the preparation of biomass samples that enable effective and accurate multi-scale microscopic analysis.
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Acknowledgements
This work was supported by the US Department of Energy Office of the Biomass Program (BSD and TBV) and the Center for direct Catalytic Conversion of Biomass to Biofuels (C3Bio), an Energy Frontier Research Center funded by the DOE, Office of Science, Office of Basic Energy Sciences under Award Number DE-SC0000997 (BSD and PNC). NREL is a national laboratory of the US Department of Energy, Office of Energy Efficiency and Renewable Energy, operated by the Alliance for Sustainable Energy, LLC.
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Donohoe, B.S., Ciesielski, P.N., Vinzant, T.B. (2012). Preservation and Preparation of Lignocellulosic Biomass Samples for Multi-scale Microscopy Analysis. In: Himmel, M. (eds) Biomass Conversion. Methods in Molecular Biology, vol 908. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-956-3_4
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DOI: https://doi.org/10.1007/978-1-61779-956-3_4
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