Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model
Transcrição
Wear sterile surgical gloves and starting from the side of incision, swab the animal with a new chlorhexidine/alcohol swab three times in a circular motion. Now, locate the incision site. Pinch the skin with tweezers, and using scissors, cut the skin layer approximately 1 centimeter from the spine below the rib cage. Make another center-cut below in the muscle layer. Locate the kidney, and without touching it, gently expose it from the abdominal cavity by applying a steady gentle pressure on the abdomen with the fingers.
Exposing the kidney is sometimes difficult. Apply gentle pressure to the abdomen and spine to avoid damaging the spleen. If organs other than the kidney come out, gently return them to the body cavity using closed forceps if necessary.
Then, gently separate the kidney from the surrounding fat just enough to visualize the renal pelvis, which can be seen as a small white dot. After visualizing the renal pelvis, use closed forceps to push down on the right side of the kidney so that the renal pelvis remains in view. Then, grasp the loaded insulin syringe in the right hand by holding it parallel to the working surface and insert the needle carefully into the renal pelvis.
Once the needle is inserted, inject the solution quickly within 3 seconds. Let the needle remain in place for approximately 10 seconds to prevent backflow of the solution. Then, remove the needle slowly and carefully and return the organ to the abdominal cavity by stretching the incision site gently using closed forceps. Now, suture the muscle layer with 5-0 absorbable sutures and place 2 to 4 independent knots. Similarly, suture the skin layer with 5-0 or 6-0 non-absorbable nylon sutures and place 2 to 4 independent knots.
