https://doi.org/10.4081/ejh.2020.3154

An improved method for preparation of mouse retinal cryosections

Vol. 64 No. 3 (2020)
Submitted: 22 June 2020
Accepted: 24 August 2020
Published: 3 September 2020
retina, cryosections, immunocytochemistry, glue
Abstract Views: 1580
PDF: 1143
Supplementary: 146
HTML: 322
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Authors

Ling Li
The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.
Yongqiong Lin
The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.
Fang Yang
The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.
Tongdan Zou
The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.
Jialiang Yang
The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.
Houbin Zhang
houbin_zhang@yahoo.com
https://orcid.org/0000-0002-2968-7454
The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.

Immunohistochemistry using mouse retinal cryosections is a routine assay used in vision research. However, retinal tissues are fragile, and it is difficult to obtain an ideal retinal cryosection. Here, we developed a modified method for preparing retinal cryosection. Super Glue was applied on the surface of the sclera before the cornea and the lens are removed from either the unfixed or PFA-fixed mouse eyeballs. The new methods largely prevented retinal detachment in mouse retinal cryosections. Immunostaining of retinal cryosections derived from PFA-fixed mouse eyes using rod and cone markers yielded high-quality immunofluorescent images. Immunolabeling of retinal cryosections obtained from unfixed mouse eyes using a cilium-specific marker had improved orientations of photoreceptor connecting cilia. This new method substantially improves the morphology and immunostaining results of fixed and unfixed mouse eyes.

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Supporting Agencies

National Natural Science Foundation of China, National Precision Medicine Project

How to Cite

Li, L., Lin, Y., Yang, F., Zou, T. ., Yang, J., & Zhang, H. (2020). An improved method for preparation of mouse retinal cryosections. European Journal of Histochemistry, 64(3). https://doi.org/10.4081/ejh.2020.3154

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