Adaptive optics retinal imaging in the living mouse eye

Area of Science:

• Ophthalmology
• Biomedical Optics
• Neuroscience

Background:

• Adaptive optics (AO) enhance in vivo mouse retinal imaging resolution.
• Previous AO attempts were limited by Shack-Hartmann wavefront sensor (SHWS) spot quality.
• Recent advances improved SHWS spot quality for mouse eye sensing.

Purpose of the Study:

• Incorporate improved SHWS techniques into a fluorescence adaptive optics scanning laser ophthalmoscope (AOSLO).
• Evaluate the performance and resolution of the developed fluorescence AOSLO on living mouse eyes.
• Demonstrate the capability for in vivo classification of retinal structures and cell types.

Main Methods:

• Integrated adjustable focus beacon with annular beam profile for improved SHWS spot quality.
• Utilized a fluorescence adaptive optics scanning laser ophthalmoscope (AOSLO) for imaging.
• Quantified transverse and axial resolutions using line spread function (LSF) and point spread function (PSF) full width at half maximum (FWHM).

Main Results:

• Obtained high-resolution images of mouse retinal structures: photoreceptor mosaic, nerve fiber bundles, capillaries, and ganglion cells.
• Achieved in vivo transverse resolution better than 0.79 μm ± 0.03 μm (45% above diffraction limit).
• Achieved in vivo axial resolution of 10.8 μm ± 0.7 μm (2x diffraction limit) with 0.36 μm axial positional accuracy.

Conclusions:

• The developed fluorescence AOSLO offers unprecedented resolution and accuracy for in vivo mouse retinal imaging.
• The instrument enables detailed visualization and classification of various retinal cell types, including bistratified ganglion cells.
• This technology advances the study of retinal structure and function in living mouse models.