0 weergaven • 2:53 min. • April 30th, 2023
- Fast an adult fish for 24 hours before injection to empty the gastrointestinal tract and create additional space in the abdomen. On the day of the procedure, anesthetize the fish with tricaine and place it into a slot in a wet sponge with the ventral side facing up.
Next, pipette the desired amount of injection solution onto a piece of laboratory film and pull it into an insulin needle. Position the needle between the pelvic fins at a 45 degree angle from the anterior-posterior body axis and gently push it in at the midline. Advance the needle approximately 1 to 2 millimeters into the abdominal cavity, the space between the abdominal wall and internal organs, and slowly inject the solution of interest. Wait for five seconds before pulling out the needle to avoid any spillage. Now, carefully remove the needle.
After injection, quickly transfer the fish into a recovery tank containing fresh tank water. If the fish doesn't begin to swim immediately, swirl the water near the gills to help it recover from anesthesia. In the example protocol, we will inject a mycobacterium marinum solution into RAG1 mutant fish to study infection.
- First, pipe out of 5 microliter droplet of the dilute
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