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Inflammation pp 367-375 | Cite as

Quantitative Assessment of Cerebral Basement Membranes Using Electron Microscopy

  • Matthew MacGregor SharpEmail author
  • Anton Page
  • Alan Morris
  • Roy O. Weller
  • Roxana O. Carare
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1559)

Abstract

In this chapter we describe in detail the tissue processing techniques we employ for the study of cerebral tissue by transmission electron microscopy (TEM). In particular, we explain a technique that enables quantification of changes in cerebral basement membranes at the ultrastructural level. This is significant, as age related pathological conditions affecting the brain are often accompanied by ultrastructural changes in the cerebral vasculature.

Briefly, experimental mice are fixed by perfusion and their brains removed. Brains are then vibratomed into 100 μm slices with regions of interest microdissected and processed for TEM following a protocol optimized for the preservation of cerebral tissue. Changes in the thickness of cerebral basement membranes are then quantified using novel software. Some prior knowledge of general TEM specimen preparation and sectioning will be useful when performing this protocol.

Key words

Transmission electron microscopy Basement membrane Tissue processing Microtomy Ultra-thin sectioning Fixation Dehydration Resin embedding Electron micrograph 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Matthew MacGregor Sharp
    • 1
    Email author
  • Anton Page
    • 2
  • Alan Morris
    • 1
  • Roy O. Weller
    • 1
  • Roxana O. Carare
    • 1
  1. 1.Clinical Neurosciences, Faculty of MedicineUniversity of SouthamptonSouthamptonUK
  2. 2.Biomedical Imaging UnitUniversity of SouthamptonSouthamptonUK

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