Application of tissue clearing and light sheet fluorescence microscopy to assess optic nerve regeneration in unsectioned tissues.
Optic nerve crush injury, as a model to study central nervous system (CNS) injury, is widely used to assess potential therapeutic strategies, aimed at promoting axon regeneration and neuronal survival. Traditional methods to evaluate optic nerve regeneration rely on histological sectioning. However, tissue sectioning results in inevitable loss of three-dimensional (3D) information, such as axonal trajectories and terminations. Here we describe a protocol for whole-tissue assessment of optic nerve regeneration in adult mice without the need for histological sectioning.
Department of Neurosurgery, Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Park Lab, LPLC, RM 3-01, 1095 NW 14th Terrace, Miami, FL, 33136, USA.
Xueting
X
Luo
Benjamin
B
Yungher
Kevin K
KK
Park
D000818:Animals; D001369:Axons / Q000473:pathology / Q000502:physiology; D004867:Equipment Design; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D009409:Nerve Crush / Q000379:methods; D009416:Nerve Regeneration; D009900:Optic Nerve / Q000502:physiology* / Q000503:physiopathology*; D020221:Optic Nerve Injuries / Q000503:physiopathology*