Abstract
The transmission electron microscope (TEM) enables a unique and valuable examination of cellular and extracellular elements in tissue in situ, in cultured cells, or in pellets derived from suspensions of cells or other materials such as nanoparticles. Here we focus on the preparation of cultured Schwann cells or Schwann cell-containing dorsal root ganglion cultures. To gain as life-like as possible views of the cellular details, it is imperative to achieve excellent preservation of the cellular structure. The steps in the preparation of cultures described in this chapter represent the results of many years of accumulated TEM images to find the best methods of preservation for Schwann cells, myelin, and basal lamina components. All the materials required are listed. The methods for fixing, dehydrating, and embedding a culture are described. Choosing an area in the culture to view, scoring it, cutting it out of the resin-embedded culture, mounting it appropriately for enface or cross-sectioning, and performing the semi-thin and thin sectioning are detailed. Explaining the way in which the sections are then stained for TEM completes the Methods section. Preservation of cultured Schwann cells and their myelin sheaths can be outstanding due to the direct and rapid but careful addition of the fixative solution to the culture dish.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bunge RP, Bunge MB, Eldridge CF (1986) Linkage between axonal ensheathment and basal lamina production by Schwann cells. Annu Rev Neurosci 9(1):305–328
Clark MB, Bunge MB (1989) Cultured Schwann cells assemble normal-appearing basal lamina only when they ensheathe axons. Dev Biol 133(2):393–404
Bates ML, Puzis R, Bunge MB (2011) Preparation of spinal cord injured tissue for light and electron microscopy including preparation for immunostaining. In: Lane E, Dunnett S (eds) Animal models of movement disorder, Neuromethods, vol 62. Springer, New York, pp 381–399
Sabatini DD, Bensch K, Barrnett RJ (1963) Cytochemistry and electron microscopy. J Cell Biol 17(1):19–58
Acknowledgments
We are grateful for the cartoons prepared by Megan Marlow at the Miami Project to Cure Paralysis. The protocols, developed over many years at the Miami Project, depended primarily upon funding from NINDS (# 09923), the Miami Project to Cure Paralysis, and the Buoniconti Fund. MBB is the Christine E. Lynn Distinguished Professor of Neuroscience. The authors declare no conflicts of interest with the contents of this chapter.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Almeida, V.W., Bates, M.L., Bunge, M.B. (2018). Preservation, Sectioning, and Staining of Schwann Cell Cultures for Transmission Electron Microscopy Analysis. In: Monje, P., Kim, H. (eds) Schwann Cells. Methods in Molecular Biology, vol 1739. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7649-2_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7649-2_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7648-5
Online ISBN: 978-1-4939-7649-2
eBook Packages: Springer Protocols