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Design-Based stereology and binary image histomorphometry in nerve assessment.

Daniel A Hunter1, Deng Pan1, Matthew D Wood1

  • 1Division of Plastic and Reconstructive Surgery, Department of Surgery, Washington University School of Medicine in Saint Louis, St Louis, MO, 63110, USA.

Journal of Neuroscience Methods
|February 20, 2020
PubMed
Summary

Stereology and histomorphometry offer comparable results for peripheral nerve analysis, with stereology using light microscopy showing higher nerve fiber area. Histomorphometry provides faster analysis but needs user input for thresholding.

Keywords:
Binary image analysisDesign-Based stereologyElectron microscopyNerve injuryNerve regenerationQuantitative histomorphometry

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Area of Science:

  • Neuroscience
  • Biomedical Engineering
  • Anatomy

Background:

  • Stereology and histomorphometry are crucial for quantifying nerve characteristics in health and disease.
  • Previous studies have not systematically compared these methods in peripheral nerves.

Purpose of the Study:

  • To investigate the reliability of design-based stereology versus semi-automated binary imaging histomorphometry for assessing healthy peripheral nerve characteristics.
  • To systematically compare stereology and histomorphometry in peripheral nerve analysis.

Main Methods:

  • Uninjured rat sciatic nerves were analyzed using both stereological analysis and histomorphometry with binary image analysis.
  • Quantification included nerve fiber number, nerve area, neural density, and fiber distribution.
  • Analyses were performed by two independent, blinded investigators using light and electron microscopy.

Main Results:

  • Both stereology and histomorphometry yielded similar results for nerve fiber number and total nerve area.
  • Stereological analysis with light microscopy reported a higher nerve fiber area compared to histomorphometry or manual measurements.
  • Electron microscopy analysis did not show this difference.

Conclusions:

  • Stereology and histomorphometry provide comparable measurements for peripheral nerve fiber number and overall nerve fiber area.
  • Stereology, particularly with light microscopy, may overestimate nerve fiber area.
  • Histomorphometry offers efficient throughput but requires manual thresholding adjustments.