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Updated: Aug 10, 2025

Live Cell Imaging of Microtubule Cytoskeleton and Micromechanical Manipulation of the Arabidopsis Shoot Apical Meristem

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Single-Cell Confinement Methods to Study Plant Cytoskeleton.

Pauline Durand-Smet¹, Antoine Chevallier², Léia Colin²

¹Laboratoire Matière et Systèmes Complexes, Unité Mixte de Recherche 7057, CNRS and Université Paris Cité, Paris cedex 13, France. pauline.durand@u-paris.fr.

Methods in Molecular Biology (Clifton, N.J.)

|February 11, 2023

Summary

This summary is machine-generated.

Researchers developed novel microwell protocols to study plant protoplast cytoskeleton dynamics. This allows observing how cell shape and mechanical stress influence microtubule and actin organization in wall-less plant cells.

Keywords:

Cytoskeleton Geometry Mechanical stress Protoplasts Single cell

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Area of Science:

Plant cell biology
Cytoskeletal dynamics
Biophysics

Background:

Cytoskeletal research in animal systems benefits from single-cell studies.
Plant cell research lacks similar systems due to the rigid cell wall, limiting studies on cytoskeleton dynamics, cell shape, and mechanical stress.

Purpose of the Study:

To present two protocols for confining wall-less plant protoplasts in microwells.
To enable the analysis of cytoskeleton dynamics in relation to cell geometry and mechanical stress.

Main Methods:

Developed two distinct microwell protocols for plant protoplasts.
Protocol 1: Agarose microwells allowing biochemical cue modification.
Protocol 2: Stiff polymer (NOA73) microwells enabling protoplast pressurization and surface coating.

Main Results:

Successfully analyzed microtubule and actin dynamics in vivo within microwells.
Demonstrated the ability to investigate the self-organization of cytoskeletal components.
Revealed the relative contributions of geometric confinement and mechanical stress to cytoskeletal organization.

Conclusions:

The presented microwell protocols overcome limitations imposed by the plant cell wall.
These methods facilitate the study of cytoskeleton dynamics under controlled geometric and mechanical conditions.
Enables a deeper understanding of how external factors influence cytoskeletal self-organization in plants.