Protocol: Pump Implantation

Species:  Mouse.

         Animals were obtained from our own mouse colony housed in our campus’ animal facility at a weight ranging 17-28 grams. 

Anesthesia used:  Ketamine / Xylazine, Isoflurane as needed.

Preparation of the surgical area: 

The surgical area that was used was aseptically sterilized, had proper lighting and contained with the appropriate surgical equipment.  This room is designated as the surgery room for all rodent surgeries.  A sterile blue pad is placed on top of a heating pad.

Preparation of surgical instruments and supplies:

All of the instruments and supplies (i.e. tools, gauze, staples, etc.) are sterilized by autoclave.  The pumps were ordered sterile.

Preparation of the animal:

         The animal was injected IP with Ketamine and maintained if necessary at % isoflurane.   Ophthalmic ointment was then placed on the animal’s eyes to prevent them from drying out during surgery.  The animal’s hair was clipped on the dorsal side vertically along the spine and horizontally from right to left flank, depending on pump size. The shaved area(s) are then prepped with betadine scrub, rubbing in a circular motion. 70% alcohol is then applied to the surgical area to remove the betadine and loose hair. This process is repeated 2 or more times. The betadine and alcohol is applied with sterile gauze.  Once the animal is prepped for surgery, it is then laid on its abdomen on a sterile blue pad with a heating pad underneath.

Preparation for the surgeon:

         The surgeon wears a facemask, hair bonnet, surgical gown and clean, sterile foot covers.  Before beginning the surgery, the surgeon thoroughly washes her hands and puts on a pair of sterile gloves.  Once the surgeon is properly gowned and gloved, she places her sterile instruments on the sterile blue pad next to the microscope as well as sterile sutures and other necessary supplies. 

Monitoring:

         After the IP Ketamine injection to the animal, it is determined fully anesthetized when it lose its reflexes.  A nose cone contained isoflurane is used only if needed during the surgical procedure.  During preparation and surgery, the anesceptic depth is determined by the animal’s respiratory pattern and by pinching the animal’s foot for reflex response.  After the post-operative procedures, the animal is placed into a clean cage, which is placed on top of a heating pad at the appropriate temperature, lined with a blue pad.  The animal is monitored until it has awakened from the procedure and is moving around the cage normally.  This usually takes 30 minutes to one hour.  The animal is then observed on a daily basis by the surgeon until sacrificed.

Analgesia: 

The animal is given a subcutaneous injection of Banamine after the procedure is finished.  The animal is also given a subcutaneous injection of room temperature sterile saline directly after the surgery is finished, prior to waking up.

Operative description: 

         With a pair of fine forceps, a 2cm horizontal incision is made approximately .5-1 cm from the right of the animal’s spine, about 3-4 cm upwards from the tail.  Separate the skin from the muscle by inserting closed, small surgical scissors between the skin and muscle and gently open the scissors as they are pulled out.  Hold onto the pump with large forceps and carefully insert the pump on top of the muscle, just under the skin layer, pushing the pump up.  Leave room for the skin to be clamped back together. Once the pump is inserted, pull the skin together with forceps and clamp the skin layers closed with sterile surgical clamps.  Once the procedure is done, the animal is placed on a covered, warm heating pad and subcutaneously injected with 1 ml of sterile saline just behind the neck.  Banamine, an analgesic, is subcutaneously injected at the right flank, .04 cc.