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Kate Grieve, Vincent Borderie, Michel Paques; Retinal imaging with full-field optical coherence microscopy. Invest. Ophthalmol. Vis. Sci. 2016;57(12):470.
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© ARVO (1962-2015); The Authors (2016-present)
To interpret full-field optical coherence microscopy (FFOCM) images of ex vivo retina.
Images of flatmounted retinas of human, primate, pig, sheep, rat, mouse and zebrafish were acquired using FFOCM. For identification of ganglion cells, samples immunolabelled against Tuj1 and Brn3a were analyzed by combining FFOCM, fluorescence confocal microscopy (FCM) and fluorescence-FFOCM. To investigate post-mortem tissue changes, time series were acquired on primate retina over 48 hours and on fresh versus fixed tissue.
With FFOCM, cell types and features such as nerve fiber bundles and RGC somas were resolved without use of contrast agents at 1µm xyz resolution. RGC somas in large mammals appeared bright with dark contours, while in rodents RGC somas appeared dark with bright contours. RGC axon to soma junctions could be traced in the 3D image stacks. Time series revealed undulation of retinal tissue samples over 48 hours, though no degradation of individual cells was detected, while paraformaldehyde fixation caused increased scattering and shrinkage.
FFOCM reveals micrometric morphological detail in the retina without the use of contrast agents. We observed interspecies differences in optical properties of RGC somas. Fixation significantly alters retinal transparency hence reducing the visibility of microscopic features.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
Rat retina in FFOCM and FCM. a cross-section; b wide field mosaic of flat-mounted section of rat retina with optic nerve located at the bottom of the image and the periphery at the top, with letters indicating locations of zoomed zones c-f; c zoom on peripheral zone with FFOCM and d similar zone in FCM with Tuj1 staining reveal axons and cells; e zoom on zone close to optic nerve with FFOCM and f with FCM with Tuj1 staining reveal axon bundles. In FFOCM, axons are bright and cells are dark centered, bright contoured. Scale bar a 100µm; b 200µm, c-f 50µm.
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