Retrograde labeling in peripheral nerve research: It is not all black and white

Ayato Hayashi, Arash Moradzadeh, Daniel A. Hunter, David H. Kawamura, Vinay K. Puppala, Thomas H.H. Tung, Susan E. Mackinnon, Terence M. Myckatyn

Research output: Contribution to journalReview articlepeer-review

43 Scopus citations

Abstract

Retrograde labeling has become an important method of evaluation for peripheral nerve regeneration after injury. We review the features of the commonly used retrograde tracers Fast Blue, Fluoro-Gold, and Fluoro Ruby in addition to the various application methods (conduit reservoir, intramuscular injection, and crystal powder application) and the techniques used to count stained neurons. Upon application of the staining techniques and dyes in a rat and mouse nerve injury model, Fluoro-Gold was found to stain the greatest number of neurons with all application methods. However, due to variability of staining intensity, neuron size, and background staining, it is difficult to count the stained neurons accurately. Fast Blue stains consistently using intramuscular injection in the mouse but fails to provide adequate staining using the muscle injection method in the rat model and shows high failure rates using the conduit reservoir technique. However, crystal dye application with Fast Blue to the cut nerve end provides excellent results. We believe that it is imperative to use the various tracers and application methods prior to their experimental use to develop a consistent standardized approach to retrograde labeling.

Original languageEnglish
Pages (from-to)381-389
Number of pages9
JournalJournal of reconstructive microsurgery
Volume23
Issue number7
DOIs
StatePublished - Oct 2007

Keywords

  • Fast blue
  • Fluoro-gold
  • Neural tracer
  • Peripheral nerve
  • Retrograde labeling

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