Avascular, neovascularised, and normally vascularised retinal areas were quantified using Image J software.
CD31 positive areas were quantified using ImageJ software and all data was expressed as means ± SD of duplicate plugs normalized to the positive control (100% vascularization).
Images were processed using Adobe PhotoShop 5.0 or 7.0 and colony areas were quantified using the Image Pro Plus 5 software.
Totally, 10 random areas were quantified using an imaging analyzing system (CUE-2, Olympus Opticals, Hamburg, Germany), and the numbers were normalized to retinal capillary area.
VEGF-stained images were taken at the lesion epicenter in axial sections at ×50, and the VEGF-positive areas were quantified using light intensity gain counting (n = 3, each).
Visceral fat areas were quantified using non-contrast CT scans (conditions: 120 kVp, 150 mA, 3 mm slice thickness, 3 mm reconstruction interval) using a Somatom Sensation 16 (Siemens, Munich, Germany).
The number of colonies/plate and colony size, expressed as average area, were quantified using RT-Image software [28].
Lesion volume, spared tissue volume, and glial scar area were quantified using GFAP staining and the Cavalieri probe at 4× in all three groups using parasagittal sections, vehicle (n = 5), hFbs (n = 5), and hCNS-SCns (n = 6).
The stained sections were scanned (2.400 dpi), the digitized images of each brain section and the infarct area were quantified using a computerized image analysis program (Image J, USA).
