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P. D'Amore, M. Saint–Geniez, C.P. Lin; Visualization of the Choroidal Circulation. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1865.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Pathophysiology of the choroidal vessels is central to the development of both wet and dry age related macular degeneration (ARMD). However, visualization of the choroidal circulation is hampered by its location beneath the retinal pigment epithelium. Thus, the goal of this work was to develop methods to reproducibly visualize choroidal vessels. Methods: The choroidal circulation was visualized by two distinct methods: Mercox vascular casts and confocal immunofluorescence microscopy. Casts of the choroidal vasculature were obtained by perfusion of Mercox resin in the entire vascular bed of 6 months old mice. Eyes are enucleated and soft tissue are digested in 20% KOH. For scanning electron microscopy, dried specimen are coated with gold palladium and scanned at 17kV. For confocal microscopy, animals were injected systemically with fluorescently–labeled antibody against PECAM–1 (CD31). Images were taken approximately 24 hours later from the outside of the globe through the intact sclera using a water immersion objective lens. Results: Using the casting technique we have obtained casts of the entire adult C57Bl/6 murine choroidal vasculature. Viewed at at low power from the posterier, major vessels can be identified including, the posterior ciliary artery, divided into the long posterior ciliary arteries and vortex vein. Observation at high power from the anterior reveals details of larger arteries and veins, and the choriocapillaris in the equatorial area. Confocal microscopy shows that the endothelial cells of the choroidal vessels as well as the capillary plexus can be stained with CD31 and imaged at different optical sectioning depths. Conclusions: Mercox vascular casts and confocal immunofluorescence microscopy both allow visualization of larger choroidal vessels and the choriocapillaris in the mouse and can be applied provide insight into normal choroidal development as well as to the pathogenesis or ARMD.
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