Purchase this article with an account.
David A. Miller, Marta Grannonico, Mingna Liu, Roman V. Kuranov, Peter A. Netland, Xiaorong Liu, Hao F. Zhang; Visible-Light Optical Coherence Tomography Fibergraphy for Quantitative Imaging of Retinal Ganglion Cell Axon Bundles. Trans. Vis. Sci. Tech. 2020;9(11):11. doi: https://doi.org/10.1167/tvst.9.11.11.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To develop a practical technique for visualizing and quantifying retinal ganglion cell (RGC) axon bundles in vivo.
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-beta-III 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.
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 semi-log 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).
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.
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.
This PDF is available to Subscribers Only