June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
AxonJ: Validation of an Automated Axon Counting Method in Rodent Models of Glaucoma
Author Affiliations & Notes
  • Kasra Zarei
    Biomedical Engineering, University of Iowa, Coralville, IA
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
  • John H Fingert
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Ophthalmology, University of Iowa, Iowa City, IA
  • Mark Christopher
    Biomedical Engineering, University of Iowa, Coralville, IA
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
  • Kathy Miller
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Ophthalmology, University of Iowa, Iowa City, IA
  • Michael G Anderson
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Ophthalmology, University of Iowa, Iowa City, IA
  • Adam Hedberg-Buenz
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA
  • Anamika Tandon
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Ophthalmology, University of Iowa, Iowa City, IA
  • Todd E Scheetz
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Ophthalmology, University of Iowa, Iowa City, IA
  • Michael David Abramoff
    Stephen A. Wynn Institute for Vision Research, Iowa City, IA
    Ophthalmology, University of Iowa, Iowa City, IA
  • Footnotes
    Commercial Relationships Kasra Zarei, None; John Fingert, None; Mark Christopher, None; Kathy Miller, None; Michael Anderson, None; Adam Hedberg-Buenz, None; Anamika Tandon, None; Todd Scheetz, None; Michael Abramoff, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 2427. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Kasra Zarei, John H Fingert, Mark Christopher, Kathy Miller, Michael G Anderson, Adam Hedberg-Buenz, Anamika Tandon, Todd E Scheetz, Michael David Abramoff; AxonJ: Validation of an Automated Axon Counting Method in Rodent Models of Glaucoma. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2427. doi: https://doi.org/.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: To validate our publicly-available, novel, automated method (AxonJ) of whole-optic nerve sections in normal and glaucomatous mouse models. Axon counts of whole nerves, though ideal for precise phenotyping, are unfeasible, hence elaborate sampling strategies are required which assume homogeneous distribution of axons across the optic nerve, while semi-automated methods lack validation in rodent models.

Methods: To maximize the strength of the validation, a variety of mouse strains were evaluated at several time points. PPD stained optic nerve sections were imaged at 40x and 100x magnification. Two human experts independently counted sampling regions at 100x magnification. AxonJ was used to determine whole nerve axon counts and axon density on all images. Manual and automated counts were compared using correlation coefficients. Extrapolated and automated 40x counts were also compared. AxonJ inter-section variability was assessed using a correlation.

Results: Counts from the AxonJ method correlated closely with counts obtained from humans at 100x, r2 = 0.95, while human-to-human correlation was r2 = 0.97. AxonJ whole nerve counts at 40x also correlated with counts extrapolated from human grading of 10% of the optic nerve area, r2 = 0.94. Inter-section variability was also low, r2 = 0.95.

Conclusions: AxonJ counts all axons in rodent whole optic nerve images with a performance equivalent to human experts counting a small subset of the axons, while inter-section repeatability is high. AxonJ is fully-automated, parameter-free, runs in less than a minute on a standard desktop, and does not make assumptions about the distribution of axons across the optic nerve. It is freely available as an ImageJ plugin.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×