Super-Resolution Imaging of Plasmodesmata Using 3D Structured Illumination Microscopy

Methods Mol Biol. 2022:2457:143-148. doi: 10.1007/978-1-0716-2132-5_8.

Abstract

Plasmodesmata (PD) have a diameter of around 30-50 nm which is well below the 200 nm limit of optical resolution, making analysis by light microscopy difficult and resolving internal structures of the PD such as the desmotubule impossible. Modern super-resolution methods such as 3D structured illumination microscopy (3D-SIM) can increase the lateral and axial resolution and work well on fixed, sectioned material. However, imaging in live plant cells requires careful optimization. Here we present a method to image PD using 3D-SIM in live BY2 cells.

Keywords: 3D-SIM; BY2; Desmotubule; Fluorescence; Plasmodesmata; Super-resolution.

MeSH terms

  • Imaging, Three-Dimensional / methods
  • Lighting*
  • Microscopy, Fluorescence / methods
  • Plant Cells
  • Plasmodesmata*