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CK Spee, E Barron, DR Hinton, SJ Ryan; A Simplified Method for the Quantitation of Laser-Induced Choroidal Neovascularization in the Rat . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1266.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To analyze the three-dimensional microvascular architecture of laser-induced choroidal neovascularization (CNV) in rats and the associated cellular behaviors associated with outer retinal wound healing. Methods: Experimental CNV was induced by laser photocoagulation-mediated disruption of Bruch’s membrane in Long Evans rats. Vessels were localized by staining with the FITC-conjugated endothelial specific Griffonia simplicifolia isolectin B4 (GS-I-B4; Vector Laboratories, Burlingame CA). Vessel staining on the whole-mount RPE-choroid-sclera complex was optically sectioned with confocal laser scanning microscopy (CLSM), (Zeiss LSM 510 software) and the digitised data set(s) were further processed and analysed with the Zeiss LSM topography program for volume quantitation. Results: This is a reproducible, simplified method that is representative of the neovascular area in laser-induced CNV. In addition, it is cost effective and requires less exhaustive hours, as does the perfusion method. Conclusion: Three-dimensional reconstruction of a CNV lesion by CLSM enabled a qualitative and quantitative analysis of the whole CNV lesion in rats. This method may contribute to the study of the cellular and molecular mechanisms of CNV and the development of new therapies against CNV.
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