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Raman Imaging of Plant Cell Walls

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The Plant Cell Wall

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2149))

Abstract

Raman imaging is a microspectroscopic approach revealing the chemistry and structure of plant cell walls in situ on the micro- and nanoscale. The method is based on the Raman effect (inelastic scattering) that takes place when monochromatic laser light interacts with matter. The scattered light conveys a change in energy that is inherent of the involved molecule vibrations. The Raman spectra are thus characteristic for the chemical structure of the molecules and can be recorded spatially ordered with a lateral resolution of about 300 nm. Based on thousands of acquired Raman spectra, images can be assessed using univariate as well as multivariate data analysis approaches. One advantage compared to staining or labeling techniques is that not only one image is obtained as a result but different components and characteristics can be displayed in several images. Furthermore, as every pixel corresponds to a Raman spectrum, which is a kind of “molecular fingerprint,” the imaging results should always be evaluated and further details revealed by analysis (e.g., band assignment) of extracted spectra. In this chapter, the basic theoretical background of the technique and instrumentation are described together with sample preparation requirements and tips for high-quality plant tissue sections and successful Raman measurements. Typical Raman spectra of the different plant cell wall components are shown as well as an exemplified analysis of Raman data acquired on the model plant Arabidopsis. Important preprocessing methods of the spectra are included as well as single component image generation (univariate) and spectral unmixing by means of multivariate approaches (e.g., vertex component analysis).

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Abbreviations

AsLS:

Asymmetric least squares

CCD:

Charge-coupled detector

CRM :

Confocal Raman microscopy

HCA:

Hierarchical cluster analysis

MCR-ALS:

Multivariate curve resolution alternating least squares

NMF:

Nonnegative matrix factorization

PCA :

Principal component analysis

PEG:

Polyethylene glycol

S/G ratio:

Syringyl–guaiacyl ratio

S/N ratio:

Signal-to-noise ratio

VCA :

Vertex component analysis

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Acknowledgments

The authors thank Martin Felhofer for support in the graphic design, Ivan Sumerskii in the group of Antje Potthast for providing the milled wood lignin reference samples and Marie-Theres Hauser for providing the Arabidopsis thaliana specimens. All people mentioned belong to the University of Natural Resources and Life Sciences, BOKU, Vienna. The work was funded by the Austrian Science Fund (FWF): START Project [Y-728-B16] and from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme grant agreement No. 681885.

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  1. Department of Nanobiotechnology, Institute of Biophysics, BOKU-University of Natural Resources and Life Sciences, Vienna, Austria

    Batirtze Prats Mateu, Peter Bock & Notburga Gierlinger

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  1. Batirtze Prats Mateu
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  2. Peter Bock
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  3. Notburga Gierlinger
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Correspondence to Notburga Gierlinger .

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  1. Botany and Plant Science School of Natural Sciences, Ryan Institute for Environmental Marine and Energy Research School of Natural Sciences, The National University of Ireland Galway, Galway, Ireland

    Zoë A. Popper

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Mateu, B.P., Bock, P., Gierlinger, N. (2020). Raman Imaging of Plant Cell Walls. In: Popper, Z. (eds) The Plant Cell Wall. Methods in Molecular Biology, vol 2149. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0621-6_15

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  • DOI: https://doi.org/10.1007/978-1-0716-0621-6_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0619-3

  • Online ISBN: 978-1-0716-0621-6

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