Cellular-resolution OCT reveals layer-specific retinal mosaics and ganglion cell degeneration in mouse retina in vivo

Cellular-resolution retinal imaging in preclinical mouse models is limited by optical aberrations and speckle noise that prevent visualization of individual cells. We developed a wavefront sensorless adaptive optics optical coherence tomography (WSAO-OCT) platform that addresses these challenges by combining real-time aberration correction with multi-volume averaging. This integrated approach enhanced image contrast by 61% and sharpness by 55%, while averaging 50 volumes effectively reduced speckle noise. Our system enabled visualization of individual cells across all major retinal layers and the retinal pigment epithelium in living mice. Immunohistochemical validation within the retinal ganglion cell (RGC) layer confirmed that 95% of optically detected cells corresponded to actual RGCs, confirming cell-type specificity. We demonstrated the method’s translational potential by tracking RGC loss following optic nerve injury. The platform detected significant cell degeneration within three days post-injury, earlier than traditional thickness-based methods, quantifying a decline from 4,407 to 1,683 cells/mm ² (62% loss) over seven days. These results establish WSAO-OCT as an effective, non-invasive tool for cellular-level investigation of retinal pathologies, suitable for longitudinal monitoring and therapeutic evaluations in preclinical models.