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

Abstract

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.

Plant phenotyping relevance

植物組織の形態、細胞壁組成、セルロース配向、リグニン化を取得するためのRaman、SHG、2PFおよび解析手法の組合せが研究の中心であり、植物表現型の画像計測法として実質的です。

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