Quantitation of ER Morphology and Dynamics

Methods Mol Biol. 2024:2772:49-75. doi: 10.1007/978-1-0716-3710-4_5.

Abstract

The plant endoplasmic reticulum forms a network of tubules connected by three-way junctions or sheet-like cisternae. Although the network is three-dimensional, in many plant cells, it is constrained to thin volume sandwiched between the vacuole and plasma membrane, effectively restricting it to a 2-D planar network. The structure of the network, and the morphology of the tubules and cisternae can be automatically extracted following intensity-independent edge-enhancement and various segmentation techniques to give an initial pixel-based skeleton, which is then converted to a graph representation. ER dynamics can be determined using optical flow techniques from computer vision or persistency analysis. Collectively, this approach yields a wealth of quantitative metrics for ER structure and can be used to describe the effects of pharmacological treatments or genetic manipulation. The software is publicly available.

Keywords: ER cisternae; Endoplasmic reticulum dynamics; Endoplasmic reticulum morphology; Lunapark; Network analysis; Persistency analysis; Phase congruency; Quantitative confocal imaging; Reticulon.

MeSH terms

  • Benchmarking*
  • Cell Membrane
  • Endoplasmic Reticulum*
  • Food
  • Plant Cells