Revised 11/25/97
Catfish are obtained from the CLAC fish room using the white buckets stored under the sink. Remove a small amount of aquarium water from the tank the fish is removed from.
In the procedure room set up instruments and pinning board.
Either transfer fish to another white bucket containing approximately 400 mls of aquarium water or remove enough water to leave behind 400 mls in the current bucket.
Prepare fixative of choice, a volume of about 100 mls is sufficient for perfusion and post fixation.
Prepare Teleost Ringers solution from prepared stocks of Part A and Part B solutions in Finger Lab.
To 100 mls of Teleost Ringers add
Note that the Xylocaine will assist in muscle relaxation and the heparin will assist in blood perfusion
Fill perfusion tube (use the smaller perfusion pump under the hood) and needle with the Ringers/drug solution removing all bubbles from the line by tapping the line thoroughly.
Place perfusion tube into the fixative solution
To anesthetize the fish add MS222 at a concentration of 1:10000 to the 400 mls of water.
The fish should cease gill movement in about 3-5 minutes indicating deep anesthesia. The fish can now be pinned well to the dissecting trays using at least 4 pins.
Expose the heart of the fish, using scissors cut a small hole into its apex. The abdomen wall of the fish can be cut to assist in blood letting.
Insert a small blunt ended needle attached to the perfusion tubing into the heart and clamp with forceps.
Start peristaltic pump and set to between 1 and 2 on the dial, be ready for rapid fish movement when fixative reaches the body of the fish.
After about 5 minutes the fish is ready for dissection.