DiI Perfusion as a Method for Vascular Visualization in Ambystoma mexicanum

Area of Science:

• Regenerative Biology
• Developmental Biology
• Vascular Biology

Background:

• Axolotl (Ambystoma mexicanum) are crucial models for regeneration research.
• Understanding vascularization during axolotl regeneration is limited.
• Existing methods for visualizing axolotl vasculature are insufficient.

Purpose of the Study:

• To establish a straightforward method for visualizing axolotl vasculature.
• To enable detailed study of revascularization patterns during axolotl regeneration.

Main Methods:

• Perfusion of 1,1'-Dioctadecy-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) dye into the axolotl aorta using a peristaltic pump.
• DiI dye, a lipophilic carbocyanine, labels endothelial cell membranes upon contact.
• Immediate post-perfusion visualization using confocal fluorescent microscopy.

Main Results:

• The DiI perfusion method successfully labels the entire vasculature of the axolotl.
• DiI fluorescence is observed in the red-orange spectrum upon excitation with a green filter.
• The procedure is efficient, taking approximately one hour for an eight-inch axolotl.

Conclusions:

• DiI perfusion offers a simple and effective way to visualize axolotl vascular networks.
• This technique is valuable for studying vascular development and regeneration in axolotls.
• The method provides insights into the dynamic process of revascularization in regenerating tissues.