Insight into plant cell wall chemistry and structure by combination of multiphoton microscopy with Raman imaging

Zsuzsanna Heiner, Ingrid Zeise, Rivka Elbaum, Janina Kneipp*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Scopus citations


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.

Original languageAmerican English
Article numbere201700164
JournalJournal of Biophotonics
Issue number4
StatePublished - Apr 2018

Bibliographical note

Publisher Copyright:
© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim


  • 2-photon autofluorescence
  • multivariate analysis
  • second harmonic imaging
  • sorghum
  • spontaneous Raman


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