InVision: An optimized tissue clearing approach for three-dimensional imaging and analysis of intact rodent eyes
Akshay Gurdita,
Philip E.B. Nickerson,
Neno T. Pokrajac,
Arturo Ortín-Martínez,
En Leh Samuel Tsai,
Lacrimioara Comanita,
Nicole E. Yan,
Parnian Dolati,
Nobuhiko Tachibana,
Zhongda C. Liu,
Joel D. Pearson,
Danian Chen,
Rod Bremner,
Valerie A. Wallace
Affiliations
Akshay Gurdita
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada
Philip E.B. Nickerson
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
Neno T. Pokrajac
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada
Arturo Ortín-Martínez
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
En Leh Samuel Tsai
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
Lacrimioara Comanita
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
Nicole E. Yan
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
Parnian Dolati
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada
Nobuhiko Tachibana
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
Zhongda C. Liu
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada
Joel D. Pearson
Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, Canada
Danian Chen
Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, Canada; Research Laboratory of Ophthalmology and Vision Sciences, State Key Laboratory of Biotherapy, Department of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
Rod Bremner
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, Canada; Department of Ophthalmology and Vision Sciences, University of Toronto, Toronto, ON M5T 3A9, Canada
Valerie A. Wallace
Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Ophthalmology and Vision Sciences, University of Toronto, Toronto, ON M5T 3A9, Canada; Corresponding author
Summary: The mouse eye is used to model central nervous system development, pathology, angiogenesis, tumorigenesis, and regenerative therapies. To facilitate the analysis of these processes, we developed an optimized tissue clearing and depigmentation protocol, termed InVision, that permits whole-eye fluorescent marker tissue imaging. We validated this method for the analysis of normal and degenerative retinal architecture, transgenic fluorescent reporter expression, immunostaining and three-dimensional volumetric (3DV) analysis of retinoblastoma and angiogenesis. We also used this method to characterize material transfer (MT), a recently described phenomenon of horizontal protein exchange that occurs between transplanted and recipient photoreceptors. 3D spatial distribution analysis of MT in transplanted retinas suggests that MT of cytoplasmic GFP between photoreceptors is mediated by short-range, proximity-dependent cellular interactions. The InVision protocol will allow investigators working across multiple cell biological disciplines to generate novel insights into the local cellular networks involved in cell biological processes in the eye.
