Aim ras

Spectral domain optical coherence tomography (SD-OCT) images were obtained using the Bioptigen Spectral Domain Ophthalmic Imaging System (SDOIS; Envisu R2200, Bioptigen, Morrisville, NC, USA). ALMS mice were anesthetized and pupils were dilated by applying 1–2 drops of topical 0.5% tropicamide (Visufarma, Rome, Italy). To prevent corneal desiccation during the procedure, topical lubricant eye drops (Recugel; Bausch & Lomb, Rochester, NY, USA) were applied bilaterally with a small brush. Mice were positioned into the animal imaging mount and rodent alignment stage (AIM-RAS; Bioptigen, Morrisville, NC, USA); the laser source was placed in front of the mouse, and images were acquired by the InVivoVue Clinic software (Bioptigen, Morrisville, NC, USA). Three images, one central, one superior, and one inferior to the optic nerve, were taken from each eye. ONL thickness was manually measured three times from each OCT scan image and averaged.

Spectral domain optical coherence tomogram images were obtained using the Bioptigen Spectral Domain Ophthalmic Imaging System (SDOIS; Envisu R2200; Bioptigen, Inc., Morrisville, NC). Mice were anesthetized, and pupils were dilated by applying one or two drops of topical 0.5% tropicamide (Visufarma, Rome, Italy). To prevent corneal desiccation during the procedure, topical lubricant eye drops (Recugel; Bausch & Lomb, Rochester, NY) were applied bilaterally with a small brush. Mice were positioned into the animal imaging mount and rodent alignment stage (AIM-RAS; Bioptigen, Inc.). The laser source was placed in front of the mouse, and images were acquired by the InVivoVue Clinic software (Bioptigen, Inc.). Three images—one central, one superior, and one inferior to the optic nerve—were taken from each eye. Outer nuclear layer thickness was manually measured three times from each OCT scan image and averaged.

Mice were anesthetized using 100 mg/kg ketamine and 10 mg/kg xylazine intraperitoneally, and the pupils were dilated using atropine drops (5 mg/mL). The mouse was placed on a rodent alignment system with a bit bar (AIM-RAS, Bioptigen, USA). The optic nerve was aligned to the center of the image on a SD-OCT imaging device with a mouse retina lens with a 50° field of view (Bioptigen Envisu R2210 VHR, Bioptigen, USA). The eyes were kept moisturized with eye drops (Systance Ultra, Alcon) and an eye gel (Vidisic carbogen, Bausch & Lomb). The following protocols (a-scan × b-scan × frame) for both eyes were run: (1) linear b-scan, 1.0 mm, 1,000 × 2 × 24 (fast fundus); (2) rectangular, 1.8 × 1.8 mm, 1,000 × 100 × 6 × 1 (high-resolution volume); (3) rectangular, 1.8 × 1.8 mm, 400 × 400 × 4 × 1 (square pixel volume); and (4) radial, 1.8 × 1.8 mm, 1,000 × 100 × 6 × 1 (high-resolution radial volume). The frames were averaged on the InVivoVue Reader software (Bioptigen, USA) and analyzed on Diver 3.4.4 software (Bioptigen, USA) for the VIP for the ONL.