Evaluation of tissue components in the peripheral nervous system using Sirius red staining and immunohistochemistry: a comparative study (human, pig, rat)

J Neurosci Methods. 2010 Jun 30;190(1):112-6. doi: 10.1016/j.jneumeth.2010.04.014. Epub 2010 Apr 22.


Little is known about species differences in the peripheral nerve system and quantitative evaluation of main tissue components has rarely been done. Nevertheless, animal models are used for example in pain research without exact knowledge of degree of fibrosis in pathological states which would determine possible treatment options. It would therefore be of crucial interest to describe the degree of fibrosis and the remaining functional nerve tissue as exact as possible. In the present study we evaluated collagen (stroma) and nerve fiber (parenchyma) composition of peripheral nerves in three species (human, rat, pig) and used digital colour-separation and analysis for collagen type differentiation and quantification of immuno-positive-stained area. We found similar ratios of collagen types I and III in epineurium and similar immuno-positive area for staining of neurofilament and S-100beta. In contrast, we measured significantly different ratios of collagen type I to type III in the endoneurium. This combined analysis of the main tissue components of peripheral nerves could be an easy-to-use tool in evaluating changes during damage caused by scaring, systemic disease or compression syndromes. The calculated collagen type I/III ratio may serve as an objective diagnostic value for the description or as prognostic marker for therapeutic approaches in peripheral nerve pathology. However, in particular studies of collagen accumulation in nerves, species dependant differences have to be considered.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Azo Compounds
  • Collagen / metabolism*
  • Collagen Type I / metabolism
  • Collagen Type III / metabolism
  • Coloring Agents
  • Histological Techniques / methods*
  • Humans
  • Immunohistochemistry / methods*
  • Male
  • Microscopy, Polarization
  • Nerve Growth Factors / metabolism
  • Neurofilament Proteins / metabolism
  • Peripheral Nerves / metabolism*
  • Rats
  • Rats, Inbred Lew
  • S100 Calcium Binding Protein beta Subunit
  • S100 Proteins / metabolism
  • Sciatic Nerve / metabolism
  • Species Specificity
  • Swine


  • Azo Compounds
  • Collagen Type I
  • Collagen Type III
  • Coloring Agents
  • Nerve Growth Factors
  • Neurofilament Proteins
  • S100 Calcium Binding Protein beta Subunit
  • S100 Proteins
  • Collagen