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 subcell...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,AOTPEFM 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 timelapse 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 highresolution 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.展开更多
基金supported by the Hong Kong Research Grants Council through grants 662513,16103215,16148816,16102518,16149316,T13-607/12R,T13-706/11-1,AOE/M-09/12,T13-605/18W,C6002-17GF,C6001-19EF and N_HKUST603/19the Hong Kong University of Science and Technology(HKUST)through grant RPC10EG33+1 种基金the Innovation and Technology Commission through grant ITCPD/17-9the Health and Medical Research Fund through grant HMRF18SC17.
文摘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,AOTPEFM 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 timelapse 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 highresolution 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.
