Invivovue diver

Retinal thickness was measured in situ in mice under anesthesia using a small animal spectral domain optical coherence tomography (SD-OCT) imaging system (Envisu R2200, Bioptigen) and InVivoVue Diver software (Bioptigen). Measurements were made at 4 compass points 350 μm from center of the optic nerve head and averaged. The total retina spans from the inner limiting membrane (ILM) to the retinal pigment epithelium (RPE). The inner retina spans from the ILM to the inner limit of the outer plexiform layer (OPL). The outer retina spans from the inner limit of the OPL to the RPE.

Spectral domain optical coherence technology/tomography (SD-OCT; Bioptigen; Durham, NC, USA), in conjunction with InVivoVue Diver software (Version 2.4) analysis, was used for in vivo assessment of retinal structure. After determining that no apparent anomalous retinal defects were present, quantitative retinal layer thickness measurements for each animal were calculated individually for the right eye (RE) and left (LE) eye of each animal. Automated total and layer-specific retinal thickness values were calculated via manual determination of 10 retinal boundaries, which defined 9 distinct retinal tissue layers. These retinal tissue layers were obtained for 8 “inner” and “outer” retinal zones (relative to the central optic nerve head (ONH)) comprising Superior, Temporal, Inferior, and Nasal positions of each retina. Mirrored retinal zone measurements for the left and right eyes of each animal were then averaged to obtain a single measurement, per animal, for each retinal zone location. Following determination of averaged individual mouse retinal layer measurements, group data were then combined to assess potential genotype-specific differences in total retinal thickness as well as for each of the smaller encapsulated subdivisions and individual retinal layers.

High-resolution spectral domain OCT was performed on the baseline and 7 days after exposure of the BALB/c mice to PSB-12489 and/or BL, as described elsewhere [33 ]. All measurements were performed under general anesthesia. The pupils of both eyes were dilated after application of 5 mg/mL solution of tropicamide. To prevent corneal drying, a Systane Ultra Eye gel (Systane^® Ultra, Norbrook, England) was applied on the cornea. The mice were fixed in the holder and ten series of 100 b-measurements were carried out (each b-measurement had 1000 a-measurements). The data obtained were aligned, averaged and a 3D image was created. Photoreceptor layer thickness was measured at 25 different points, which were selected using InVivoVueDiver (Bioptigen, JAV) software. The central point was targeted at the center of the optic nerve. The photoreceptor layer thickness was estimated by measuring the distance between the outer plexiform layer and the external limiting membrane.