Visible-light optical coherence tomography fibergraphy for quantitative imaging of retinal ganglion cell axon bundles

David A. Miller, Marta Grannonico, Mingna Liu, Roman V. Kuranov, Peter A. Netland, Xiaorong Liu*, Hao F. Zhang

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Purpose: To develop a practical technique for visualizing and quantifying retinal ganglion cell (RGC) axon bundles in vivo. Methods: We applied visible-light optical coherence tomography (vis-OCT) to image the RGC axon bundles, referred to as vis-OCT fibergraphy, of healthy wild-type C57BL/6 mice. After vis-OCT imaging, retinas were flat-mounted, immunostained with anti-betaIII tubulin (Tuj1) antibody for RGC axons, and imaged with confocal microscopy. We quantitatively compared the RGC axon bundle networks imaged by in vivo vis-OCT and ex vivo confocal microscopy using semi-log Sholl analysis. Results: Side-by-side comparison of ex vivo confocal microscopy and in vivo vis-OCT confirmed that vis-OCT fibergraphy captures true RGC axon bundle networks. The semilog Sholl regression coefficients extracted from vis-OCT fibergrams (3.7 ± 0.8 mm–1)and confocal microscopy (3.6 ± 0.3 mm–1) images also showed good agreement with each other (n = 6). Conclusions: We demonstrated the feasibility of using vis-OCT fibergraphy to visualize RGC axon bundles. Further applying Sholl analysis has the potential to identify biomarkers for non-invasively assessing RGC health. Translational Relevance: Our novel technique for visualizing and quantifying RGC axon bundles in vivo provides a potential measurement tool for diagnosing and tracking the progression of optic neuropathies.

Original languageEnglish (US)
Article number11
Pages (from-to)1-12
Number of pages12
JournalTranslational Vision Science and Technology
Issue number11
StatePublished - 2020


  • Ganglion cell axon bundles
  • Image analysis
  • Optical coherence tomography

ASJC Scopus subject areas

  • Biomedical Engineering
  • Ophthalmology


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