Preparation of spinal cord injured tissue for light and electron microscopy including preparation for immunostaining

Margaret L. Bates, Raisa Puzis, Mary Bartlett Bunge

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

An important outcome measure of effects of treatment for experimental spinal cord injury is excellent histology. One way to achieve this is to prepare the tissue for electron microscopy, which ensures the best preservation and requires resin embedding. "Semi-thin" sections of resin-embedded tissue (0.6-1.5 μm in thickness) may be stained and viewed in the light microscope to detect morphological features of the lesion such as various cell types, myelin and blood vessels. Specific areas of interest then may be chosen for thin sectioning preparatory to examination in the electron microscope when better resolution is required, such as searching for non-myelinated nerve fibers. The steps required to prepare the tissue for both light and electron microscopy are detailed in this chapter. A method for assessing the total numbers of axons, myelinated and non-myelinated, is included. Immunostaining for both light and electron microscopy also is described.

Original languageEnglish (US)
Title of host publicationAnimal Models of Movement Disorders
Subtitle of host publicationVolume II
EditorsEmma Lane, Stephen Dunnett
Pages381-399
Number of pages19
DOIs
StatePublished - Oct 24 2011

Publication series

NameNeuromethods
Volume62
ISSN (Print)0893-2336
ISSN (Electronic)1940-6045

Keywords

  • Green fluorescent protein label
  • Myelinated axons
  • Non-myelinated axons
  • Resin embedding
  • Schwann cells
  • Spinal cord injury
  • Tissue preservation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Neuroscience(all)
  • Psychiatry and Mental health

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