Skip to main content

Use of Confocal Microscopy for Three-Dimensional Imaging of Neurons in the Spinal Cord

  • Protocol
  • First Online:
Book cover Histology Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 611))

  • 4790 Accesses

Abstract

Confocal microscopy provides a powerful and efficient tool for studying the morphology of cells. Here we describe its use to study the morphology of neurons in the dorsal horn of the spinal cord either following electrophysiological studies in live tissue slices or in neurons filled with dye in fixed tissue sections following identification using retrograde tracing. The methods are broadly applicable to other cell types and can be combined with multiple label immunohistochemistry to study cellular constituents or with subsequent DAB staining to produce a permanent mount.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Balasubramanyan, S., Stemkowski, P.L., Stebbing, M.J., and Smith, P.A. (2006) Sciatic chronic constriction injury produces cell-type-specific changes in the electrophysiological properties of rat substantia gelatinosa neurons. J. Neurophysiol. 96, 579–590.

    Article  PubMed  Google Scholar 

  2. Furness, J.B., Robbins, H.L., Xiao, J., Stebbing, M.J., and Nurgali, K. (2004) Projections and chemistry of Dogiel type II neurons in the mouse colon. Cell Tissue Res. 317, 1–12.

    Article  PubMed  CAS  Google Scholar 

  3. Young, H.M., Kunze, W.A., Pompolo, S., Furness, J.B., and Bornstein, J.C. (1994) Combined intracellular injection of Neurobiotin and pre-embedding immunocytochemistry using silver-intensified gold probes in myenteric neurons. J. Neurosci. Methods 51, 39–45.

    Article  PubMed  CAS  Google Scholar 

  4. Buhl, E.H. and Lubke, J. (1989) Intracellular lucifer yellow injection in fixed brain slices combined with retrograde tracing, light and electron microscopy. Neuroscience 28, 3–16.

    Article  PubMed  CAS  Google Scholar 

  5. Chery, N. and de Koninck, Y. (1999) Junctional versus extrajunctional glycine and GABA(A) receptor-mediated IPSCs in identified lamina I neurons of the adult rat spinal cord. J. Neurosci. 19, 7342–7355.

    PubMed  CAS  Google Scholar 

  6. Sakmann, B. and Stuart, G. (1995) Patch-pipette recordings from the soma, dendrites, and axon of neurons in brain slices. In Single-Channel Recording, Sakmann, B. and Neher, E. (Eds). Plenum Press, New York.

    Chapter  Google Scholar 

  7. Kunze, W.A., Furness, J.B., and Bornstein, J.C. (1993) Simultaneous intracellular recordings from enteric neurons reveal that myenteric AH neurons transmit via slow excitatory postsynaptic potentials. Neuroscience 55, 685–694.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Stebbing, M., Potocnik, S., Ye, P., Badoer, E. (2010). Use of Confocal Microscopy for Three-Dimensional Imaging of Neurons in the Spinal Cord. In: Hewitson, T., Darby, I. (eds) Histology Protocols. Methods in Molecular Biology, vol 611. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-345-9_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-345-9_14

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-344-2

  • Online ISBN: 978-1-60327-345-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics