Preparation of Onion Epidermal Cell Walls for Imaging by Atomic Force Microscopy (AFM).

Abstract

The growing plant cell wall is comprised of long, thin cellulose microfibrils embedded in a hydrated matrix of polysaccharides and glycoproteins. These components are typically constructed in layers (lamellae) on the inner surface of the cell wall, i.e ., between the existing wall and the plasma membrane. The organization of these components is an important feature for plant cell growth and mechanics. To directly visualize the nano-scale structure of the newly-deposited surface of primary plant cell walls without dehydration or chemical extraction, a protocol of cell wall preparation for AFM imaging the most recently-synthesized cell wall surface in aqueous solutions was developed. Although the method was developed for onion scale epidermal peels, it can also be adapted to other organs, such as Arabidopsis hypocotyls, as well as ground samples of cell walls from the leaf petioles or hypocotyls of Arabidopsis and cucumber, maize coleoptiles and onion parenchyma. Potential artifacts of AFM imaging of plant cell walls are also discussed.

Plant phenotyping relevance

植物細胞壁のナノスケール構造をAFMで直接可視化するための試料調製プロトコルを開発しており、植物形態・構造の取得法が中心である。

Code and data availability

This is a Bio-protocol methods article describing onion epidermal cell wall preparation for AFM imaging. No public phenotype/trait datasets, AFM image data deposits, author analysis code, or trained models are mentioned; analysis is done in proprietary Nanoscope software, and no repository or availability URL appears.

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