PETIT, Jules; GLAVIER, Marie; BROCARD, Lysiane; BAYER, Emmanuelle

doi:10.1007/978-1-0716-2132-5_3

Métadonnées

Afficher la notice complète

Licence d’utilisation du document

PETIT, Jules

Laboratoire de biogenèse membranaire [LBM]

GLAVIER, Marie

Laboratoire de biogenèse membranaire [LBM]

BROCARD, Lysiane

Langue

Article de revue

Ce document a été publié dans

Methods in Molecular Biology. 2022-03-30, vol. 2457, p. 57-74

Résumé en anglais

Plant plasmodesmata (PD) are complex intercellular channels consisting of a thin endoplasmic reticulum (ER) tubule enveloped by the plasma membrane (PM). PD were first observed by electron microscopy about 50 years ago and, since, numerous studies in transmission and scanning electron microscopy have provided important information regarding their overall organization, revealing at the same time their diversity in terms of structure and morphology. However, and despite the fact that PD cell-cell communication is of critical importance for plant growth, development, cellular patterning, and response to biotic and abiotic stresses, linking their structural organization to their functional state has been proven difficult. This is in part due to their small size (20-50 nm in diameter) and the difficulty to resolve these structures in three dimensions at nanometer resolution to provide details of their internal organization.In this protocol, we provide in detail a complete process to produce high-resolution transmission electron tomograms of PD. We describe the preparation of the plant sample using high-pressure cryofixation and cryo-substitution. We also describe how to prepare filmed grids and how to cut and collect the sections using an ultramicrotome. We explain how to acquire a tilt series and how to reconstruct a tomogram from it using the IMOD software. We also give a few guidelines on segmentation of the reconstructed tomogram.< Réduire

Mots clés en anglais

Electron Microscope Tomography

Microscopy

Electron

Scanning

Microtomy

Plant Cells

Plasmodesmata

Métadonnées

Partager cette publication !

Licence d’utilisation du document

Plasmodesmata Ultrastructure Determination Using Electron Tomography.

Langue

Ce document a été publié dans

Résumé en anglais

Mots clés en anglais

URI

DOI

Unités de recherche