Mydriacil

To perform fundus examination, mice were anaesthetized by an intraperitoneal injection of ketamine (80 mg/kg) and xylazine (10 mg/kg). The pupils were then dilated with 0.5% tropicamide drops (Mydriacil; Alcon, Geneva, Switzerland), and corneas were kept moist with GenTeal eye gel (Novartis, Basel, Switzerland). Fundus imaging was performed using a Micron III retinal imaging system (Phoenix Research Laboratories, Pleasanton, CA, USA) with StreamPix 5 software. Brightness and contrast adjustments were uniformly applied to images in Adobe Photoshop. Immediately after in vivo imaging, mice were euthanized for tissue collection.

Mice were clinically examined on day (d) 0, 14, 21, 28 and 35 post-immunization (p.i.). Mice were anesthetized by an i.p. injection of a mixture of ketamine (80 mg/kg; Troy Laboratories, Glendenning, NSW, Australia) and xylazine (10 mg/kg; Troy Laboratories). The pupils were dilated with 0.5% tropicamide (Mydriacil, Alcon Laboratories, Vilvoorde, Belgium), and the cornea was kept moist with the application of sterile lubricant GenTeal gel (Novartis, North Ryde, NSW, Australia). The fundus was examined using the Micron III camera (Phoenix Research Laboratories, Pleasanton, CA, USA) with StreamPix 5 software. Examination consisted firstly of capturing short 5 seconds videos with 100 frames in brightfield, followed by a sequence of GFP⁺ or YFP⁺ cells using the green fluorescent barrier filters, and lastly by a short sequence following a 20 μl s.c. injection of 10% fluorescein isothiocyanate (Alcon Laboratories, Frenchs Forest, NSW, Australia) to visualize the retinal vasculature. The brightness and contrast of all fundus images were adjusted equally using ImageJ 1.48C software [27 ].