Retinal vasculature was stained and leukostasis was analyzed as previously described [15 (link),23 (link)]. Saline was perfused into the aorta to clear non-adherent leukocytes, then 10 mL of fluorescein-labeled concanavalin A lectin (1 mg/mL in PBS; Vector laboratories Burlington, ON Canada) was perfused to stain the retinal vasculature. After enucleation and isolation of the retina, flat mounts were imaged using a fluorescent stereoscope and the number of leukocytes adhered to the vasculature wall were counted (n = 3 samples/group). The 500 μm and 50 μm scale bar displayed gives a visual indicator of the size of the representative image.
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