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

doi:10.1016/j.isci.2021.102905

InVision: An optimized tissue clearing approach for three-dimensional imaging and analysis of intact rodent eyes

iScience. 2021 Jul 26;24(8):102905. doi: 10.1016/j.isci.2021.102905. eCollection 2021 Aug 20.

Authors

Akshay Gurdita^{1

2}, Philip E B Nickerson¹, Neno T Pokrajac^{1

2}, Arturo Ortín-Martínez¹, En Leh Samuel Tsai¹, Lacrimioara Comanita¹, Nicole E Yan¹, Parnian Dolati^{1

2}, Nobuhiko Tachibana¹, Zhongda C Liu¹, Joel D Pearson³, Danian Chen^{3

4}, Rod Bremner^{2

3

5}, Valerie A Wallace^{1

2

5}

Affiliations

¹ Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, 60 Leonard Avenue, Toronto, ON M5T 2S8, Canada.
² Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, Canada.
³ Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, Canada.
⁴ Research Laboratory of Ophthalmology and Vision Sciences, State Key Laboratory of Biotherapy, Department of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China.
⁵ Department of Ophthalmology and Vision Sciences, University of Toronto, Toronto, ON M5T 3A9, Canada.

Abstract

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.

Keywords: Biological sciences; Biological sciences research methodologies; Biology experimental methods; Biotechnology; Neuroscience; Optical imaging.