A New Method to Visualize Retinal Deep Vascular Layers Using Whole-Mount Tissue Preparations

The mouse retina is a widely used model to study sprouting angiogenesis. Most blood vessel staining protocols focus on the superficial vascular layer. Here, we describe a method to obtain thick sections of retina whole-mounts that allow for detailed visualization of diving vascular sprouts, and deep and intermediate-layer angiogenesis. This protocol does not require extra tissue processing and can be coupled with any previously optimized protocol for retina whole-mount staining.