Fig. 4

Regrown nerve resembles the contralateral uninjured nerve in 3D spatial positioning, resulting in functional vision restoration. A) Depth-coded imaging at day 14 PL (left side regrown). B) Orthogonal plane view with nerves intersecting the brain at day 14 PL. C) Overview of 3D volume colocalization analysis using Imaris image analysis software. Regrown nerve side is labelled in green and contralateral nerve side is pseudo-colored red. C’) The two halves of the tadpole head are overlayed on the mirror plane in 3D space C”) Each optic nerve is made into a “surface” from which the overlap volume ratio can be calculated. D) The overlapped volume ratio refers to a measurement that calculates the proportion of volume shared between two 3D surface objects, in this case, the overlayed optic nerves. Results of the single cut overlay, double cut overlay and naïve overlay cohorts are shown. There were no statistical differences at day 14 between single cut, double cut or naïve cohorts. E) Tadpole behavioral vision test setup. In summary, the setup features a nested tank design, with an inner tank containing the tadpole and water, while the outer tank is divided into black and white halves. Every two minutes, the outer tank is rotated. A camera mounted above the nested tanks captures footage of the tadpole as it swims. F) Quantification of percentage of time tadpoles swam in the light side before injury (naïve) and at Day 3, 7, 14 and 21 PL. G) Results of following the same individual tadpoles across these timepoints. T1 = tadpole 1.