Xenopus Oocyte Harvest

Xenopus oocyte harvest is a performed when the milking of frogs is not possible and carries the potential to damage oocytes. Therefore, surgery is necessary for the removal and collection of the oocytes.

Procedure Preparation

1. As per Yale IACUC Policy Transportation of Animals (Intra), animals are transported to the room where the procedure will be performed.

2. The work space will be organized with all tools needed to minimize manipulation of the animal.

Procedure

3. Frog will be anesthetized to alleviate potential pain associated with the procedure. Refer to the Anesthesia Policy, Guidelines and Procedure document for recommending anesthetics.

4. The frog is placed in dorsal recumbency. Gross debris will be washed away, and the surgical site should be rinsed clean using sterile saline.

5. Artificial slime (such as (Shieldex ™ or Aquatronic) or conditioned water will be applied to the frog’s skin to keep it moist, taking care not to introduce conditioned water into the surgical site.  Moistening or irrigation of this site will be performed with sterile saline as necessary.   

6. A diagonal skin incision, approximately 1cm, is made in a lower quadrant of the abdomen. Sides will be alternated to allow maximum healing time.

7. The skin is gently tented using sterile surgical forceps, and an incision is made through the muscle layer to expose the ovary.

8. Using forceps, the ovary will be grasped, and the oocyte masses exteriorized.

9. A mass of oocytes is excised, and the remainder will be carefully placed back into the coelomic cavity.

10.The muscle and skin incisions will be closed using appropriately sized suture material.

Post-Procedure Monitoring and Care

11.Animal is placed on moistened paper towels until mobile, or in a shallow tray of room temperature conditioned water until able to swim.

12.Animals should be checked daily for at least 3 days for any signs of wound dehiscence and inflammation.

13.If any evidence of clinical disease occurs, VCS will be contacted.