A label-free, fast and high-specificity technique for plant cell wall imaging and composition analysis

Abstract

Abstract Background New cell wall imaging tools permit direct visualization of the molecular architecture of cell walls and provide detailed chemical information on wall polymers, which will aid efforts to use these polymers in multiple applications; however, detailed imaging and quantification of the native composition and architecture in the cell wall remains challenging. Results Here, we describe a label-free imaging technology, coherent Raman scattering (CRS) microscopy, including coherent anti-Stokes Raman scattering (CARS) microscopy and stimulated Raman scattering (SRS) microscopy, which can be used to visualize the major structures and chemical composition of plant cell walls. We outline the major steps of the procedure, including sample preparation, setting the mapping parameters, analysis of spectral data, and image generation. Applying this rapid approach will help researchers understand the highly heterogeneous structures and organization of plant cell walls. Conclusions This method can potentially be incorporated into label-free microanalyses of plant cell wall chemical composition based on the in situ vibrations of molecules.

Code and data availability

This is a methods/protocol paper on CRS microscopy of plant cell walls. The only URLs mentioned are generic tools (Fiji/ImageJ, microscope control software) and the article's own DOI and license links. No public phenotype/trait datasets, plant images, author analysis code, trained models, or supplements containing them

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