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bioRxiv · 10.64898/2026.03.21.712744

Computational aberration-corrected volumetric imaging of single retinal cells in the living eye

Feng, G.·Godinez, D. R.·Li, Z.·Nolen, S.·Cho, H.·Kimball, E.·Duh, E. J.·Johnson, T. V.·Yi, J.
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Abstract

The eye offers a unique non-invasive window for accessing single-cell level structures and functions of the central nervous system (CNS) throughout the retina. However, strong and space-varying ocular aberrations, along with limited volume rates, challenge large-scale cellular imaging in living eyes and stymie the full potential of possible biological and pathological studies in retina. Here, we present plenoptic illumination scanning laser ophthalmoscopy (PI-SLO), a 3D fluorescent retinal imaging modality that enables high-speed, widefield, volumetric single-cell imaging with low phototoxicity. By capturing multiple angular images of fluorescence signals from the entire volume, PI-SLO enables digital aberration correction and 3D imaging across a >20{o} FOV with >23 Hz volume rate. We leverage this structural and functional imaging modality to investigate three key aspects of CNS physiology through the living mouse retina, including: microglial process dynamics, vascular perfusion, and light evoked calcium fluxes in inner retinal neurons. PI-SLO is a versatile non-invasive platform for in vivo investigation of retinal and CNS physiology at the cellular level.

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Feng, G., Godinez, D. R., Li, Z., Nolen, S., Cho, H., Kimball, E., Duh, E. J., Johnson, T. V., Yi, J.. 2026-03-24. Computational aberration-corrected volumetric imaging of single retinal cells in the living eye. https://doi.org/10.64898/2026.03.21.712744

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