Ultrastructural Analysis and Quantification of Peroxisome-Organelle Contacts

Methods Mol Biol. 2023:2643:105-122. doi: 10.1007/978-1-0716-3048-8_8.

Abstract

Transmission electron microscopy (TEM) has long been a vital technology to visualize the interaction of cellular compartments at the highest possible resolution. While this paved the way to describing organelles within the cellular context in detail, TEM has long been underused to generate quantitative data, analyzing those interactions as well as underlying mechanisms leading to their formation and modification. Here we describe a simple stereological method to unbiasedly assess the extent of organelle-organelle membrane contact sites, able to efficiently generate accurate and reproducible quantitative data from cultured mammalian cells prepared for TEM.

Keywords: Cell culture; Electron microscopy; Membrane contact site; Peroxisomes; Quantitation; Stereology; Transmission electron microscopy.

MeSH terms

  • Animals
  • Cells, Cultured
  • Mammals
  • Microscopy, Electron, Transmission
  • Organelles* / ultrastructure
  • Peroxisomes*