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Insight into plant cell wall chemistry and structure by combination of multiphoton microscopy with Raman imaging
Author(s) -
Heiner Zsuzsanna,
Zeise Ingrid,
Elbaum Rivka,
Kneipp Janina
Publication year - 2018
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201700164
Subject(s) - raman spectroscopy , microscopy , autofluorescence , two photon excitation microscopy , raman scattering , chemistry , raman microscope , biophysics , second harmonic generation , chemical imaging , hyperspectral imaging , cell wall , materials science , fluorescence , optics , laser , biology , physics , remote sensing , geology , biochemistry
Spontaneous Raman scattering microspectroscopy, second harmonic generation (SHG) and 2‐photon excited fluorescence (2PF) were used in combination to characterize the morphology together with the chemical composition of the cell wall in native plant tissues. As the data obtained with unstained sections of Sorghum bicolor root and leaf tissues illustrate, nonresonant as well as pre‐resonant Raman microscopy in combination with hyperspectral analysis reveals details about the distribution and composition of the major cell wall constituents. Multivariate analysis of the Raman data allows separation of different tissue regions, specifically the endodermis, xylem and lumen. The orientation of cellulose microfibrils is obtained from polarization‐resolved SHG signals. Furthermore, 2‐photon autofluorescence images can be used to image lignification. The combined compositional, morphological and orientational information in the proposed coupling of SHG, Raman imaging and 2PF presents an extension of existing vibrational microspectroscopic imaging and multiphoton microscopic approaches not only for plant tissues.