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

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

STAR Protoc

Wellcome-Wolfson Institute for Experimental Medicine, Queen's University, Belfast BT9 7BL, UK. Electronic address:

Published: December 2024

AI Article Synopsis

Article Abstract

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.

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Source
http://dx.doi.org/10.1016/j.xpro.2024.103510DOI Listing

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