Protocol for Detecting mRNAs in Mouse Retinal Vasculature
Author Information
Author(s): Augustine Josy, Smith Madeleine R., Delaney Ryan, Owuamalam Precious O., Costa Guilherme
Primary Institution: Queen’s University, Belfast
Hypothesis
Can single-molecule fluorescent in situ hybridization (smFISH) be effectively used to detect individual mRNAs in the developing mouse retinal vasculature?
Conclusion
The protocol allows for high-resolution detection of individual mRNAs in the retinal vasculature of post-natal mice.
Supporting Evidence
- The protocol enables the detection of individual mRNAs in vascular endothelial cells.
- It provides simple steps to overcome challenges related to tissue permeabilization.
- The method allows for mRNA and protein co-detection in the developing retinal vasculature.
- smFISH offers high spatial resolution for transcript detection.
Takeaway
This study shows a way to see tiny pieces of genetic material in the blood vessels of baby mice's eyes, helping scientists understand how these vessels grow.
Methodology
The protocol involves preparing mouse retinas, performing smFISH to detect mRNAs, and using imaging techniques to visualize the results.
Limitations
The protocol is limited to a couple of exemplar transcripts and does not provide a strategy for endothelial cell segmentation.
Participant Demographics
C57BL/6J, post-natal 3–20 days, either gender
Digital Object Identifier (DOI)
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