Retinal microvascular and neuronal pathologies probedin vivoby adaptive optical two-photon fluorescence microscopy

Abstract

The retina, behind the transparent optics of the eye, is the only neural tissue whose physiology and pathology can be non-invasively probed by optical microscopy. The aberrations intrinsic to the mouse eye, however, prevent high-resolution investigation of retinal structure and functionin vivo. Optimizing the design of a two-photon fluorescence microscope (2PFM) and sample preparation procedure, we found that adaptive optics (AO), by measuring and correcting ocular aberrations, is essential for resolving synapses and achieving three-dimensional cellular resolution in the mouse retinain vivo. Applying AO-2PFM to longitudinal retinal imaging in transgenic models of retinal pathology, we characterized microvascular lesions and observed microglial migration in a proliferative vascular retinopathy model, and found Lidocaine to effectively suppress retinal ganglion cell hyperactivity in a retinal degeneration model. Tracking structural and functional changes at high resolution longitudinally, AO-2PFM enables microscopic investigations of retinal pathology and pharmacology for disease diagnosis and treatmentin vivo.

In Situ Imaging of Retinal Calcium Dynamics in Awake Animals

“…In the past decade, multiple groups demonstrated the exquisite capability of imaging retinal structures in vivo using two-photon (2P) microscopy with adaptive optics (Geng et al, 2012, Qin et al, 2020, Biss et al, 2007, Wahl et al, 2016, Yin et al, 2013, Zhang et al, 2022, including reports of functional imaging of light response (Qin et al, 2020, Yin et al, 2013, Zhang et al, 2022. However, setting up and operating an imaging system with adaptive optics is non-trivial for most research groups.…”