A Method to Visualize Cell Proliferation of Arabidopsis thaliana : A Case Study of the Root Apical Meristem.

Abstract

Plant growth and development rely on a delicate balance between cell proliferation and cell differentiation. The root apical meristem (RAM) of Arabidopsis thaliana is an excellent model to study the cell cycle due to the coordinated relationship between nucleus shape and cell size at each stage, allowing for precise estimation of the cell cycle duration. In this study, we present a method for high-resolution visualization of RAM cells. This is the first protocol that allows for simultaneous high-resolution imaging of cellular and nuclear stains, being compatible with DNA replication markers such as EdU, including fluorescent proteins (H2B::YFP), SYTOX DNA stains, and the cell wall stain SR2200. This protocol includes a clarification procedure that enables the acquisition of high-resolution 3D images, suitable for detailed subsequent analysis.

Code and data availability

The paper describes a triple-staining/3D imaging protocol with Cellpose/Plant-Seg segmentation, but no public phenotype datasets, images, code, or trained models are deposited; only supplementary protocol files (Data S1–S4, Movies S1–S5) are mentioned without any public repository or URL. Allowed URLs are only the CC-B

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