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Transmission Electron Microscopy and Morphometry of the CNS White Matter

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Axon Degeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2143))

Abstract

Transmission electron microscopy of central nervous system white matter has provided unparalleled access to the ultrastructural features of axons, their myelin sheaths, and the major cells of white matter; namely, oligodendrocytes, oligodendrocyte precursors, astrocytes, and microglia. In particular, it has been invaluable in elucidating pathological changes in axons and myelin following experimentally induced injury or genetic alteration, in animal models. While also of value in the examination of human white matter, the tissue is rarely fixed adequately for the types of detailed analyses that can be performed on well-preserved samples from animal models, perfusion fixed at the time of death. In this chapter we describe methods for obtaining, processing, and visualizing white matter samples using transmission electron microscopy of perfusion fixed tissue and for unbiased morphometry of white matter, with particular emphasis on axon and myelin pathology. Several advanced electron microscopy techniques are now available, but this method remains the most expedient and accessible for routine ultrastructural examination and morphometry.

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Acknowledgments

J.M.E. is grateful to Professor Ian Griffiths who developed the morphometric methods described herein, and in whose lab she received training in morphometry of CNS white matter. The fixation and processing methods were developed and optimized over many years by Professor Ian Griffiths and Mrs. Mailis McCulloch in the Applied Neurobiology Group at the University of Glasgow. Mrs. McCulloch and Mrs. Jennifer Barrie provided excellent technical assistance for many of the studies cited in this chapter. Past and present EM work in J.M.E.’s lab is funded by the Multiple Sclerosis Society, UK. I.D.D. is grateful to Ben August for his EM expertise and Abigail Radcliff for her help with preparation of tissues. His work has been supported by many grants including the NMSS grant # RG 1501-02876 and the Boespflug Foundation. Colin Crawford assisted with photography for Fig. 1.

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Authors and Affiliations

  1. Institute of Infection, Immunity and Inflammation, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow, UK

    Julia M. Edgar & Rebecca Sherrard Smith

  2. Department of Neurogenetics, Max Planck Institute for Experimental Medicine, Goettingen, Germany

    Julia M. Edgar

  3. Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, USA

    Ian D. Duncan

Authors
  1. Julia M. Edgar
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  2. Rebecca Sherrard Smith
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  3. Ian D. Duncan
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Corresponding author

Correspondence to Julia M. Edgar .

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Editors and Affiliations

  1. Department of Biochemistry, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, NY, USA

    Elisabetta Babetto

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Edgar, J.M., Smith, R.S., Duncan, I.D. (2020). Transmission Electron Microscopy and Morphometry of the CNS White Matter. In: Babetto, E. (eds) Axon Degeneration. Methods in Molecular Biology, vol 2143. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0585-1_18

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  • DOI: https://doi.org/10.1007/978-1-0716-0585-1_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0584-4

  • Online ISBN: 978-1-0716-0585-1

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