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Ch 8.Immunohistoch / immunology
Ch 10.GC/MS, NMR and Proteomics
Tail Vein Injection Techniques (1) (2)
Species: Mouse. (weight ranging
17-28 )grams.
Anesthesia used: None
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./procedures. A sterile blue pad is laid on top of a heating pad.
Preparation of surgical
instruments and supplies: All of the instruments and supplies are sterilized by autoclave. The needles and syringes are ordered
sterile packaged.
Preparation of the animal: The animal is placed into a clear, plastic mouse restraint so that the animal
is not freely mobile, but its tail is able to be handled. Its tail and tail only is place into warm water (approximately 40oC)
so that the tail vein is more visible. Once the tail is heated to adequate temperature, the animal is ready for the procedure.
A heat lamp may be used if the water is not adequately warming the tail for the injection.
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:
The animal is monitored during the procedure by observing its respiratory rate and making sure that the animal is in any distress
while being in the restraint. The restraint does not harm the animal. While the animal’s tail is warming in the water,
careful precautions are taken so that the animal does not get wet with water other than its tail. The animal is not submersed
in water. After the procedure is done the animal is monitored to make sure it is not stressed and returns back to its normal
respiratory rate, if elevated.
Analgesia: No analgesic is needed for this procedure.
Operative description: Once
the animal’s tail is warmed to adequate temperature then it is ready for the procedure. The animal, still in the restraint,
is placed on the sterile blue pad. The tail is wiped with alcohol to sterilize the area to be injected. A 30 gauge needle
is used with a syringe of a total volume of 100ul to inject. With your left hand, hold onto the tail so that the vein is facing
upward, facing you. While holding the syringe in your right hand (if right-handed), carefully inject the needle into the vein,
while still holding onto the vein so that it does not roll. Inject the solution slowly so that it flows at a slow pace through
the vein. You will see the tail color turn pale at first. Once the injection is done, the mouse is released from the restraint
and put back in its cage and observed.
Protocol 4. INTRAVENOUS (IV) INJECTION
MOUSE (Tail Vein): Restrain the mouse with
physical or chemical restraint. Rotate the tail slightly to visualize vein. Disinfect injection site and insert needle (27-30 gauge)
into the vein at a slight angle. You will not be able to aspirate, instead inject slowly and watch for clearing of the lumen. Incorrect
positioning will result in a slight bulge in the tail. If this occurs, remove needle and repeat process proximal to previous site.
Upon completion remove needle and apply pressure to injection site.
RAT (Tail or Saphenous Vein): Tail injection procedure same as
in the mouse. For the saphenous vein, restrain the rat with the use of anesthesia. Extend the hind leg and shave hair to expose lateral
saphenous vein. Disinfect the injection site and apply tourniquet-like pressure to the upper portion of the leg. Insert needle into
the vein and aspirate. Release tourniquet pressure and inject. Upon completion remove needle and ensure proper hemostasis. The recommended
needle size for IV injections in the rat is 22-25 gauge.