Harvesting an Inguinal Draining Lymph Node from a Murine Model

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Mice used for the experiment

1. Use 8-week-old male and female mice on the B6 background.

2. Surgery procedure to remove the inguinal draining lymph node

1. Euthanize mice using CO₂ asphyxiation followed by cervical dislocation.
2. Immobilize mice on the acrylic stage with adhesive tape and apply mineral oil with a cotton swab to the abdominal skin to prevent fur deposition around the incision (Figure 1B). Fur removal is not necessary.
3. Perform a midline incision using microsurgery curved scissors (11.5 cm) and microsurgery curved forceps (12.5 cm) in the abdomen from the pubis to the xiphoid process (Figure 1C).
4. Dissociate the abdominal musculature from the skin (Figure 1D).
5. Make horizontal skin incisions at the top and bottom of the vertical incision line to create skin flaps on the side of interest (according to the side of the antibody mix injection) and flap the skin to visualize the lymph node (Figure 1E).
6. Tape the skin flap on the acrylic plate (Figure 1F).
7. Remove the inguinal draining lymph node using microsurgery curved forceps (Figure 1F). Lymph nodes will appear as translucid, usually bilobular, spheres under the skin.