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Marie Darche, Morgane Belle, Stéphane Fouquet, Alain Chédotal, Ilaria Cascone, Michel Paques; 3D visualization of angiogenesis in the mouse eye using light sheet fluorescence microscopy. Invest. Ophthalmol. Vis. Sci. 2019;60(9):198.
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During development, physiological angiogenesis occurs through a succession of vascular growth and regression. In the eye, several layers successively develop : the tunica vasculosa lentis (TVL), the vasa hyaloidea propria (VHP), both forming the hyaloid vascular system (HVS), and the retinal vessels. The HVS ultimately regresses as the retinal vasculature develops. When this dynamic is disturbed, various abnormalities such as persisting HVS and/or defective retinal vascularization may occur and impair visual function.Understanding ocular vessel development ideally requires simultaneous evaluation of the networks, which is cumbersome by classical histology. Indeed, section and dissection of ocular tissues lead to partial destruction of their integrity and a total loss of their spatial organization.
In this work, we used a combination of light-sheet fluorescence microscopy (LSFM) and iDISCO+ clearing tissue preparation (adapted from Renier et al. 2016) on albino mice’s intact eyes. The acquired serial images were then reconstructed in 3D and the networks analyzed using Imaris software.
We used this approach to image in 3D the entire intact vascular networks and perivascular cells in the developing eye (from embryo to adult mice) and during oxygen-induced retinopathy.
We show that LSFM and iDISCO+ clearing combination is a powerful tool to study vascular networks in the intact eye. Contrary to traditional imaging techniques, this method is rapid, non-destructive and can be used to analyze large tridimensional networks in their natural spatial organization. This method will provide new insights to the exploration of physiological and pathological ocular angiogenesis.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
3D visualization of hyaloid vessels in the mouse pup’s eye (P4 and P9).
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