Adaptive optics two-photon microscopy enables near-diffraction-limited and functional retinal imaging in vivo
Open Access
- 6 May 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Light: Science & Applications
- Vol. 9 (1), 1-11
- https://doi.org/10.1038/s41377-020-0317-9
Abstract
In vivo fundus imaging offers non-invasive access to neuron structures and biochemical processes in the retina. However, optical aberrations of the eye degrade the imaging resolution and prevent visualization of subcellular retinal structures. We developed an adaptive optics two-photon excitation fluorescence microscopy (AO-TPEFM) system to correct ocular aberrations based on a nonlinear fluorescent guide star and achieved subcellular resolution for in vivo fluorescence imaging of the mouse retina. With accurate wavefront sensing and rapid aberration correction, AO-TPEFM permits structural and functional imaging of the mouse retina with submicron resolution. Specifically, simultaneous functional calcium imaging of neuronal somas and dendrites was demonstrated. Moreover, the time-lapse morphological alteration and dynamics of microglia were characterized in a mouse model of retinal disorder. In addition, precise laser axotomy was achieved, and degeneration of retinal nerve fibres was studied. This high-resolution AO-TPEFM is a promising tool for non-invasive retinal imaging and can facilitate the understanding of a variety of eye diseases as well as neurodegenerative disorders in the central nervous system.Keywords
Funding Information
- Research Grants Council, University Grants Committee (16149316)
- Food and Health Bureau of the Government of the Hong Kong Special Administrative Region | Health and Medical Research Fund (HMRF18SC17)
This publication has 84 references indexed in Scilit:
- Structural and Functional Abnormalities of Retinal Ganglion Cells Measured In Vivo at the Onset of Optic Nerve Head Surface Change in Experimental GlaucomaInvestigative Ophthalmology & Visual Science, 2012
- Retinal Ganglion Cell Morphology after Optic Nerve Crush and Experimental GlaucomaInvestigative Ophthalmology & Visual Science, 2012
- Changes in physiological properties of rat ganglion cells during retinal degenerationJournal of Neurophysiology, 2011
- Optical properties of the mouse eyeBiomedical Optics Express, 2011
- Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopyBiomedical Optics Express, 2010
- Adaptive Optics Retinal Imaging: Emerging Clinical ApplicationsOptometry and Vision Science, 2010
- Rodent models of glaucomaBrain Research Bulletin, 2010
- In Vivo Imaging of Microscopic Structures in the Rat RetinaInvestigative Ophthalmology & Visual Science, 2009
- In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell MosaicInvestigative Ophthalmology & Visual Science, 2009
- A pyramid approach to subpixel registration based on intensityIEEE Transactions on Image Processing, 1998