Preparation and Imaging of Specialized ER Using Super-Resolution and TEM Techniques

Karen Bell; Karl Oparka; Kirsten Knox

doi:10.1007/978-1-0716-3710-4_4

Preparation and Imaging of Specialized ER Using Super-Resolution and TEM Techniques

Methods Mol Biol. 2024:2772:39-48. doi: 10.1007/978-1-0716-3710-4_4.

Authors

Karen Bell¹, Karl Oparka¹, Kirsten Knox²

Affiliations

¹ Institute of Molecular Plant Sciences, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
² School of Molecular Biosciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK. Kirsten.Knox@glasgow.ac.uk.

PMID: 38411805
DOI: 10.1007/978-1-0716-3710-4_4

Abstract

The plant endoplasmic reticulum (ER) forms several specialized structures. These include the sieve element reticulum (SER) and the desmotubule formed as the ER passes through plasmodesmata. Imaging both of these structures has been inhibited by the resolution limits of light microscopy and their relatively inaccessible locations, combined with the fragile nature of the ER. Here we describe methods to view desmotubules in live cells under 3D-structured illumination microscopy (3D-SIM) and methods to fix and prepare phloem tissue for both 3D-SIM and transmission electron microscopy (TEM), which preserve the fragile structure and allow the detailed imaging of the SER.

Keywords: 3D-SIM; BY2; Desmotubule; Endoplasmic reticulum; Imaging; OMX; Sieve-element reticulum; ZIO.

MeSH terms

Endoplasmic Reticulum*
Microscopy, Electron, Transmission
Phloem*
Plasmodesmata