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Parenteral Injections
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Procedure:-

 

Mouse Handling and Manual Restraint:

 

 

  1. Remove the mouse from the cage by gently grasping the tail at the base. Place the animal on a wire-bar cage lid to permit grasping. (Avoid exciting the mouse with loud noises or sharp movement. Slight, rearward traction on the tail will result in instinctive grasping on the wire-bar cage lid. )
  2. Approach the back of the neck from the rear with the free hand. Firmly grasp the skin behind the ears with the thumb and index finger.
  3. Transfer the tail from preferred hand to beneath the little finger of the hand holding the scruff of the neck.
  4. Observe or inject the restrained mouse.

 

"Restraining technique can still vary depending on the injection methodology."

 

Types of Parenteral Injections:

 

a) Intramuscular Injection:

 

Materials Required:

 

  • Injectate
  • 1- to 3-ml syringe with 22- to 30-G needle

 

Procedure:

 

  1. Fill syringe with injectate and remove air bubbles.
  2. Manually restrain as described before.
  3. Before inserting the needle in the heavy musculature of the upper thigh, rub the area with an Alcohol swab.
  4. After injecting Injectate, withdraw the needle and pressurize the area with a cotton ball for a while and place the animal back.

 

b) Intradermal Injection:

 

Materials Required:

 

  • 70% ethanol.
  • Injectate.
  • Clippers with #40 blade.
  • Gauze sponge or swab.
  • 1- to 3-ml syringe with 25- to 30-G needle.
  • Additional reagents and equipments for handling and restraint and anesthesia.

 

Procedure:

 

  1. Restrain and anesthetize the animal.
  2. Use clippers to remove hair from an area on the back and wipe with 70% ethanol on a gauze sponge or swab.
  3. Fill syringe with injectate and remove air bubbles.
  4. Pull the skin taut with one hand and insert the needle, bevel up and at a shallow angle, just under the superficial layer of epidermis.
  5. Inject 50 μl for the mouse. (A bleb will appear at the injection site when needle placement is correct.

 

c) Subcutaneous Injection:

 

Materials Required:

 

  • Injectate.
  • 1- to 3-ml syringe with 22- to 30-G needle.

 

Procedure:

 

  1. Fill syringe with injectate and remove air bubbles.
  2. Manually restrain the mouse.
  3. Insert the needle under the skin of the interscapular area tented by the restraining thumb and forefinger.
  4. Inject with moderate pressure and speed.

 

d) Intravenous Injection:

 

Materials Required:

 

  • Injectate.
  • 70% ethanol.
  • 1-ml syringe with 25- to 30-G needle.
  • Restrainer.
  • Heat lamp or beaker containing warm water.
  • Gauze sponge or swab.

 

Procedure:

 

  1. Fill syringe with injectate and remove air bubbles. (Removal of air bubbles is critical to avoid air embolism.)
  2. Place mouse in a restrainer. Warm the tail with a heat lamp or by immersing in warm water to dilate vessels.
  3. Swab the tail with 70% ethanol on a gauze sponge or swab.
  4. Immobilize the tail with gentle traction.
  5. Visualize the lateral tail vein and insert needle parallel to the vein 2 to 4 mm into the lumen. Keep the bevel of the needle facing up.
  6. Inject slowly. (No bleb should form if needle was properly located. If a bleb appears, indicating failure to cannulate the vein, additional attempts may be made proximally. Thus it is helpful to make the first attempt at injection as close to the tip of the tail as possible.)
  7. Withdraw the needle and apply digital pressure if necessary to achieve hemostasis.

 

e) Intraperitoneal Injection:

 

Materials Required:

 

  • Injectate.
  • 1- or 3-ml syringe with 22- to 25-G needle.

 

Procedure:

 

  1. Manually restrain the animal.
  2. For the mouse, rat, or hamster, expose the animal's abdomen, tilting the head downward. (This procedure is usually done with one person restraining the animal while the other injects the substance.)
  3. Fill syringe with injectate and remove air bubbles.
  4. Insert needle into the lower left or right quadrant of the abdomen, avoiding the abdominal midline.
  5. Inject with moderate pressure and speed.

 

f) Footpad Injection:

 

Materials Required:

 

  • Injectate.
  • 1-ml syringe with 25-G needle.

 

Procedure:

 

  1. Manually restrain the animal. (This procedure is usually done with one person restraining the animal while the other injects the substance.)
  2. Spread the toes of one hind foot and introduce the needle into the soft tissue pad of the plantar surface.(Alternatively, the injection can be made subcutaneously between the second and third digits.)
  3. Inject no more than 50 μl. Apply direct pressure following withdrawal of needle.

 

g) Intrathymic Injection:

 

Materials Required:

 

  • 70% ethanol.
  • Injectate.
  • Surgical board.
  • Gauze sponge or swab.
  • Scalpel handle (#3) and scalpel blade (#15).
  • Thumb forceps.
  • Iris scissors, straight.
  • 1-ml syringe with 28- to 30-G needle.
  • Wound clips or 4-0 suture material with needle.
  • Additional reagents and equipment for anesthesia.

 

Procedure:

 

  1. Anesthetize the mouse and immobilize in dorsal recumbency on a surgical board.
  2. Swab the chest area with 70% ethanol on a gauze sponge or swab.
  3. Make a 1-cm midline incision with scalpel and blade at the thoracic inlet,i.e at the junction of the lower cervical and upper thoracic regions.
  4. Use thumb forceps and iris scissors to longitudinally bisect the upper one-third of the sternum and expose the thymus.
  5. Fill syringe with injectate and remove air bubbles.
  6. Insert needle into the parenchyma of the thymus and inject up to 10 μl in either lobe.
  7. Withdraw needle and close wound with wound clips or 4-0 suture material.

 

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