Ultrastructural analysis of adult mouse neocortex comparing aldehyde perfusion with cryo fixation

Elife. 2015 Aug 11:4:e05793. doi: 10.7554/eLife.05793.

Abstract

Analysis of brain ultrastructure using electron microscopy typically relies on chemical fixation. However, this is known to cause significant tissue distortion including a reduction in the extracellular space. Cryo fixation is thought to give a truer representation of biological structures, and here we use rapid, high-pressure freezing on adult mouse neocortex to quantify the extent to which these two fixation methods differ in terms of their preservation of the different cellular compartments, and the arrangement of membranes at the synapse and around blood vessels. As well as preserving a physiological extracellular space, cryo fixation reveals larger numbers of docked synaptic vesicles, a smaller glial volume, and a less intimate glial coverage of synapses and blood vessels compared to chemical fixation. The ultrastructure of mouse neocortex therefore differs significantly comparing cryo and chemical fixation conditions.

Keywords: astrocyte; axon; dendrite; mouse; neuroscience; synapse; ultrastructure.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehydes / metabolism
  • Animals
  • Fixatives / metabolism
  • Freezing
  • Mice
  • Microscopy, Electron / methods
  • Neocortex / ultrastructure*
  • Perfusion
  • Tissue Fixation / methods*

Substances

  • Aldehydes
  • Fixatives

Grants and funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.