High resolution fundus imaging by confocal scanning laser ophthalmoscopy in the mouse

We evaluated fundus imaging using a modified confocal scanning laser ophthalmoscope (cSLO) in mice. Examinations were performed in conscious, untrained mice. The largest field of view measured 1520 × 1520 μ, with a significant interindividual variability, itself correlated to biometric variability. The composite field of view extended up to the ora serrata. The reflectance imaging associated light reflection from nerve fiber bundles and vessel walls, and absorption by hemoglobin and melanin. Light absorption by the pigment epithelium indeed increased the contrast of the nerve fiber layer, but impaired viewing of the choroid. Due to the confocal mode, fluorescence angiograms with clear separation of retinal and choroidal fluorescence could be obtained even in albino mice. Micrometric-scale transverse resolution and several planes of optical sectioning within the retina were obtained. This permitted for instance tridimensional, subcellular viewing of gfp-expressing retinal microglial cells in CX3CR1 mice. We concluded that cSLO is a promising tool for noninvasive, multimodal intravital microscopy of the fundus in the mouse.