Visualization of Neural and Vascular Networks in a Chicken Embryo

Begin with a windowed, fertilized chicken egg containing an early-stage embryo with its neural tube excised at the level of somites one to seven.

Take a GFP-labeled donor neural tube from a stage-matched transgenic chicken embryo and implant it into the recipient, ensuring proper anatomical orientation.

Remove excess liquid around the graft to enhance adhesion between the donor and host tissues.

Seal the window with clear tape to prevent dehydration and contamination.

Incubate the egg until the embryo develops to the desired stage.

Remove the tape.

Identify a vitelline vein on the yolk that carries blood toward the embryo and remove the overlying vitelline membrane.

Using a glass needle, steadily inject a lipophilic fluorescent dye into the vein.

Visualize the embryo under a stereofluorescence microscope.

The injected dye binds to the endothelial cells lining the blood vessels, highlighting the vascular network, while the GFP-labeled neural tube enables neural network tracking.

When ready to begin the grafting procedure, carefully transfer the dissected neural tube from the watch glass to the host embryo. Position the neural tube in the correct orientation. And gently push the explant adjac