Protocol for single-molecule FISH in the developing mouse retinal vasculature.

Autor:	Augustine J; Wellcome-Wolfson Institute for Experimental Medicine, Queen's University, Belfast BT9 7BL, UK. Electronic address: j.augustine@qub.ac.uk., Smith MR; Wellcome-Wolfson Institute for Experimental Medicine, Queen's University, Belfast BT9 7BL, UK., Delaney R; Advanced Imaging and Histology Unit, Faculty of Medicine, Health and Life Sciences, Queen's University, Belfast BT9 7BL, UK., Owuamalam PO; Wellcome-Wolfson Institute for Experimental Medicine, Queen's University, Belfast BT9 7BL, UK., Costa G; Wellcome-Wolfson Institute for Experimental Medicine, Queen's University, Belfast BT9 7BL, UK. Electronic address: g.costa@qub.ac.uk.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2024 Dec 20; Vol. 5 (4), pp. 103510. Date of Electronic Publication: 2024 Dec 12.
DOI:	10.1016/j.xpro.2024.103510
Abstrakt:	The developing vasculature of the post-natal mouse retina is a powerful model to discover mechanisms of vessel formation and to test modulators of neovascularization. We present a protocol for single-molecule fluorescent in situ hybridization (smFISH) in whole-mount mouse retinas enabling the detection of individual mRNAs in vascular endothelial cells. We describe procedures from initial retina preparation to smFISH and detection. Our approach offers simple steps to overcome challenges related to tissue permeabilization, mRNA and protein co-detection, and post-acquisition image processing. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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