apte r s niederkorn j y isolation and characterization of a unique natural killer cell inhibitory factor present in the anterior chamber of the eye j immunol 156 2667 2673 1996 briefly mice were anesthetized by injecting ketamine hydrochloride 100 mg kg and xylazine 13 4 mg kg intraperitoneally and their pupils were dilated with 1 tropicamide using argon green laser four laser burns were placed around the optic nerve 0 1 second 50 u03bcm and 110 mw seven days after laser mice were anesthetized as described above and perfused intraventricularly with fitc dextran sigma aldrich mice were euthanized in a co2 chamber and their eyes were harvested for tissue processing a dissecting microscope was used to remove the cornea and lens and to gently separate the retina from the underlying choroid and sclera micro scissors were used to make four radial incisions in the sclerochoroidal u201c eyecup u201d to prepare choroidal flat mounts on glass slides the tissues were incubated in 4 paraformaldehyde for 45 min and washed 3 times with 3 bovine serum albumin the choroidal flat mounts were analyzed for the presence of cnv by confocal microscopy the extent of choroidal neovascularization was quantified by metamorph imaging software.