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Each nerve was analyzed over a length of 200 µm by measuring the area spanned by the clogs compared to the total area, employing the analysis package included in Image Pro Plus (Media Cybernetics, Silver Springs, MD).
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Calibrated SD-OCT scans measuring 6 X 6 X 2 mm centered on the fovea and the optic nerve were analyzed in 50 healthy right eyes.
Five midsagittal sections containing the optic nerve (or sections within 32 µm of the optic nerve) were analyzed in three non-overlapping microscope fields, according to their eccentricity ("central", "middle" and "peripheral").
Ten non-serial sections, except sections containing the optic nerve, were analyzed per eye.
For each animal, three fields of view at × 20 magnification in central retinal areas (in proximity to the optic nerve) were analyzed and at least three sections were quantified.
Transverse semi-thin sections at the injury sites of sciatic nerves were analyzed by Toluidine blue staining.
The longitudinal sections at the injury sites of sciatic nerves were analyzed by hematoxylin and eosin (H.E).
For each genotype, at least four nerves were analyzed (22-day old Dicerfl/fl Dhh-Cre+ and control Dicerwt/fl Dhh-Cre+ littermates).
Nerves were analyzed at 3 days post-injection.
At 4 weeks post-vector administration, animals were killed and the brain, spinal cord and sciatic nerves were analyzed by fluorescent microscopy.
In randomly selected high power fields, the total number of nerve fibers, myelin sheath thickness and myelinated fiber diameter of the regenerated nerves were analyzed with Image Pro Plus 6.0.
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