Imaging the adult zebrafish cone mosaic using optical coherence tomography

Huckenpahler, A.L., Wilk, M.A., Cooper, R.F., Moehring, F., Link, B.A., Carroll, J., Collery, R.F.
Visual neuroscience   33: E011 (Journal)
Registered Authors
Collery, Ross, Link, Brian
Cone mosaic, Optical coherence tomography, Retina, Zebrafish
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Green Fluorescent Proteins/genetics
  • Microscopy, Fluorescence
  • Retinal Cone Photoreceptor Cells/cytology*
  • Tomography, Optical Coherence*
  • Zebrafish/anatomy & histology*
  • Zebrafish/genetics
28177275 Full text @ Vis. Neurosci.
Zebrafish (Danio rerio) provide many advantages as a model organism for studying ocular disease and development, and there is great interest in the ability to non-invasively assess their photoreceptor mosaic. Despite recent applications of scanning light ophthalmoscopy, fundus photography, and gonioscopy to in vivo imaging of the adult zebrafish eye, current techniques either lack accurate scaling information (limiting quantitative analyses) or require euthanizing the fish (precluding longitudinal analyses). Here we describe improved methods for imaging the adult zebrafish retina using spectral domain optical coherence tomography (OCT). Transgenic fli1:eGFP zebrafish were imaged using the Bioptigen Envisu R2200 broadband source OCT with a 12-mm telecentric probe to measure axial length and a mouse retina probe to acquire retinal volume scans subtending 1.2 × 1.2 mm nominally. En face summed volume projections were generated from the volume scans using custom software that allows the user to create contours tailored to specific retinal layer(s) of interest. Following imaging, the eyes were dissected for ex vivo fluorescence microscopy, and measurements of blood vessel branch points were compared to those made from the en face OCT images to determine the OCT lateral scale as a function of axial length. Using this scaling model, we imaged the photoreceptor layer of five wild-type zebrafish and quantified the density and packing geometry of the UV cone submosaic. Our in vivo cone density measurements agreed with measurements from previously published histology values. The method presented here allows accurate, quantitative assessment of cone structure in vivo and will be useful for longitudinal studies of the zebrafish cone mosaics.
Genes / Markers
Figure Gallery
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Engineered Foreign Genes
Errata and Notes