MethodsX · 9 authors, 3 centres
This summary was generated by AI from a single paper. It has not been reviewed by a clinician and is not clinical advice. Verify against the source before acting on it.
This paper provides a step-by-step protocol for performing corneal thermal cauterization in mice to induce and study inflammation-driven angiogenesis and lymphangiogenesis. The protocol covers thermal cauterization, cornea enucleation and dissection, immunofluorescence staining of neovasculature, and morphometric analysis, aiming to improve reproducibility across research groups.
The assay involves thermal injury of the transparent, avascular mouse cornea, which induces polymorphonuclear leukocyte recruitment and subsequent blood and lymphatic vessel formation from the limbal area. The protocol details procedures for thermal cauterization, enucleation and dissection of corneas, immunofluorescence staining (using CD31 for blood endothelial cells and LYVE-1 for lymphatic endothelial cells), and morphometric analysis of vessel area and branch point density. The authors note that thermal cauterization is less expensive and less time-consuming than alternative corneal injury models, though it demands skillful technique due to the small size of the mouse eye. The protocol discusses ethical considerations, alternative endothelial markers, and potential applications including inhibitor blockade, siRNA knockdown, viral vector delivery, and knockout mouse models.
